Lab Data: Interpretation Factors

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Questions and Answers

What is the primary purpose of the color-coding system used for blood collection tubes?

  • To denote the expiration date of the tube's contents
  • To easily identify the type of anticoagulant or additive present in the tube (correct)
  • To differentiate between tubes used for serum and plasma collection
  • To indicate the volume of blood the tube can hold

Serum is best described as the liquid component of blood:

  • Separated from blood cells in a heparinized tube.
  • Obtained after centrifuging a tube with anticoagulant.
  • Remaining after blood has clotted and the clot is removed. (correct)
  • Collected in a tube containing EDTA.

For a routine mammalian Complete Blood Count (CBC), which blood collection tube is typically the tube of choice?

  • Green top (Heparin) tube
  • Blue top tube
  • Red top tube
  • Purple top (EDTA) tube (correct)

Why is it important to avoid filling an EDTA tube before a red top tube when collecting multiple blood samples?

<p>To minimize the risk of anticoagulant carryover from the EDTA tube into the red top tube. (B)</p> Signup and view all the answers

Which of the following is considered a pre-analytical source of error in laboratory testing?

<p>Hemolysis of the blood sample due to improper collection technique (C)</p> Signup and view all the answers

Over- or under-filling a blood collection tube can lead to inaccurate results primarily due to:

<p>Alteration of the anticoagulant-to-blood ratio. (C)</p> Signup and view all the answers

In diagnostic testing, sensitivity is defined as the:

<p>Proportion of truly positive cases correctly identified by the test. (B)</p> Signup and view all the answers

What is the potential consequence of using a small vein for blood collection compared to a large vein?

<p>Increased likelihood of sample dilution with tissue fluid. (A)</p> Signup and view all the answers

Prolonged draw time during blood collection should be avoided because it can lead to:

<p>Activation and clumping of platelets. (C)</p> Signup and view all the answers

Using an incorrect anticoagulant for a specific blood test is an example of which type of error?

<p>Pre-analytical error (B)</p> Signup and view all the answers

If blood is allowed to clot in a tube intended for plasma collection, which component of the blood will be missing or significantly reduced in the resulting sample?

<p>Fibrinogen (D)</p> Signup and view all the answers

Which color-topped vacutainer tube is specifically designed for coagulation tests?

<p>Blue top (Citrate) (C)</p> Signup and view all the answers

For glucose determinations, particularly when glycolysis inhibition is required, which vacutainer tube is most appropriate?

<p>Gray top (Potassium Oxalate/Sodium Fluoride) tube (A)</p> Signup and view all the answers

Hemolysis, a pre-analytical error, can falsely elevate the results of which of the following analytes?

<p>Aspartate transaminase (AST) (D)</p> Signup and view all the answers

Lipemia in a blood sample, characterized by turbidity, can interfere with laboratory assays and falsely increase the measurement of:

<p>Hemoglobin (C)</p> Signup and view all the answers

What is the primary reason for avoiding forcing a blood sample through the needle into a collection tube?

<p>To avoid hemolysis caused by mechanical force. (B)</p> Signup and view all the answers

Incompletely filled EDTA tubes can lead to red blood cell crenation due to:

<p>Excess EDTA causing osmotic effects. (D)</p> Signup and view all the answers

Expired reagents are a source of which type of laboratory error?

<p>Analytical error (A)</p> Signup and view all the answers

What is the role of calibrators in laboratory assays?

<p>To configure the instrument to produce accurate results by using samples with known values. (B)</p> Signup and view all the answers

Laboratory controls are used primarily to:

<p>Verify the consistency and reproducibility of assay results by evaluating reagent recovery. (C)</p> Signup and view all the answers

Reporting results to the wrong patient is an example of which category of laboratory error?

<p>Post-analytical error (C)</p> Signup and view all the answers

Why is it emphasized that 'bad data is worse than no data' in the context of laboratory results?

<p>Because incorrect results can lead to inappropriate and potentially harmful clinical decisions. (D)</p> Signup and view all the answers

Chemistry results from dehydrated patients may be higher in concentrated plasma due to:

<p>Hemoconcentration leading to a higher proportion of analytes per unit volume. (A)</p> Signup and view all the answers

Enzyme activities in a blood sample, measured in U/L, can be affected by:

<p>Both the amount of enzyme and the time elapsed since sample collection. (A)</p> Signup and view all the answers

Reference intervals for laboratory tests are ideally established based on:

<p>Data from approximately 120 or more healthy individuals. (C)</p> Signup and view all the answers

Why should each laboratory ideally establish its own reference intervals?

<p>Because reference values are dependent on methodology, reagents, and patient population. (C)</p> Signup and view all the answers

In a box and whisker plot of platelet counts, outliers are identified and should be excluded primarily because they:

<p>May be caused by interfering factors like platelet clumps, leading to inaccurate representation of the population. (D)</p> Signup and view all the answers

If a laboratory has fewer than 20 healthy animals to establish a reference interval, what is the recommended alternative?

<p>Present the mean value and histogram of the observed data instead of a reference interval. (B)</p> Signup and view all the answers

A reference interval is designed to reflect approximately what percentage of a healthy population?

<p>95% (D)</p> Signup and view all the answers

A diagnostic cut-off, as an alternative to a reference interval, is primarily established by:

<p>Setting an assay value that best differentiates between diseased and non-diseased animals, often focusing on animals with the disease of interest and other diseases. (A)</p> Signup and view all the answers

Increased alkaline phosphatase (ALP), calcium (Ca), and phosphorus (Phos) levels might be considered within the reference range for:

<p>Growing animals (D)</p> Signup and view all the answers

A twofold increase in serum potassium (K) is generally considered more clinically significant than a twofold increase in alanine aminotransferase (ALT) because:

<p>Potassium plays a more critical role in vital functions, and even small changes can be life-threatening. (A)</p> Signup and view all the answers

It is possible for sick animals to have test results within the reference interval, especially in early stages of disease. For example, a normal creatinine level can be seen in:

<p>Early renal disease (B)</p> Signup and view all the answers

Which of the following assay diagnostic properties is most crucial for a screening test?

<p>High sensitivity (A)</p> Signup and view all the answers

A diagnostic test with high specificity is best suited for:

<p>Confirming a diagnosis after a positive screening test. (D)</p> Signup and view all the answers

If a test has 95% sensitivity, what does this imply regarding false negative results?

<p>There is a 5% chance of a false negative. (B)</p> Signup and view all the answers

In the context of predictive values, disease prevalence significantly affects:

<p>Positive and negative predictive values (PPV and NPV). (D)</p> Signup and view all the answers

Calculate the accuracy of a diagnostic test given the following values: True Positives (TP) = 6, True Negatives (TN) = 966, False Positives (FP) = 7, False Negatives (FN) = 21.

<p>97.2% (D)</p> Signup and view all the answers

What is the primary focus of the initial part of the lecture on laboratory tests?

<p>General principles influencing lab test results and their interpretation. (B)</p> Signup and view all the answers

Which category of laboratory errors is most directly controlled by the veterinarian?

<p>Pre-analytical errors related to sample collection and handling. (A)</p> Signup and view all the answers

If a veterinarian sends a sample to a reference lab, which type of error are they LEAST likely to have direct control over?

<p>Analytical errors. (A)</p> Signup and view all the answers

What is a potential consequence of using a small vein for blood collection that might NOT occur with a larger vein?

<p>Vein collapse and hemolysis due to negative pressure. (A)</p> Signup and view all the answers

Why is a prolonged blood draw time more likely to cause platelet clumps in a sample?

<p>Extended exposure of platelets to the syringe and needle, promoting aggregation. (D)</p> Signup and view all the answers

In blood collection, vacutainers are advantageous in human medicine primarily because they:

<p>Directly transfer blood into tubes containing additives, reducing clot formation time. (D)</p> Signup and view all the answers

Choosing the correct blood collection tube is crucial. What could be a consequence of using the incorrect tube?

<p>Sample clotting when anticoagulants are needed, or using the wrong anticoagulant. (B)</p> Signup and view all the answers

Serum differs from plasma primarily due to the:

<p>Inclusion of clotting factors in plasma but not in serum. (A)</p> Signup and view all the answers

What is the function of the gel in a serum separator tube (SST)?

<p>To create a physical barrier between serum and blood cells after centrifugation. (B)</p> Signup and view all the answers

Why is it important to allow a red top tube to fully clot before centrifugation?

<p>To ensure complete consumption of clotting factors and proper serum separation. (A)</p> Signup and view all the answers

For which of the following analytes would a red top tube be MOST appropriate?

<p>Serum glucose. (A)</p> Signup and view all the answers

Why are serum separator tubes (SST) generally NOT recommended for therapeutic drug monitoring, such as phenobarbital levels?

<p>The gel in SST tubes can absorb certain drugs, leading to falsely low results. (B)</p> Signup and view all the answers

EDTA prevents blood coagulation by:

<p>Chelating calcium ions, which are essential for the coagulation cascade. (B)</p> Signup and view all the answers

A chemistry profile result shows a critically low calcium level (e.g., 2 mg/dL) from a sample collected in a purple top tube. What is the MOST likely explanation?

<p>Contamination of the sample with EDTA from the tube. (D)</p> Signup and view all the answers

Why is a purple top (EDTA) tube considered a good choice for cell counts?

<p>EDTA preserves the morphology of cells and prevents clotting, keeping cells free for accurate counting. (A)</p> Signup and view all the answers

Lithium heparin is preferred over sodium heparin for general chemistry profiles because:

<p>Lithium does not interfere with electrolyte measurements, unlike sodium. (A)</p> Signup and view all the answers

For 'stat' (immediate) chemistry testing, which anticoagulant tube type is MOST advantageous and why?

<p>Green top tube (heparin), because it does not require clot formation time. (D)</p> Signup and view all the answers

In a situation where a very small blood volume is obtained (e.g., from a small animal), which tube type is most versatile if both CBC and chemistry are needed?

<p>Green top tube (heparin), allowing for both hematology and chemistry from a single sample. (C)</p> Signup and view all the answers

Sodium citrate (blue top tubes) is used for coagulation testing because it:

<p>Chelates calcium weakly, allowing for controlled coagulation when calcium is reintroduced. (B)</p> Signup and view all the answers

Gray top tubes containing potassium oxalate and sodium fluoride are primarily used for:

<p>Glucose determinations, especially when glycolysis inhibition is needed. (C)</p> Signup and view all the answers

If a glucose sample is not processed promptly and is left at room temperature, what is the expected change in glucose levels and why?

<p>Glucose levels will decrease as blood cells metabolize glucose (glycolysis). (B)</p> Signup and view all the answers

How does refrigeration help in preserving glucose levels in a blood sample?

<p>Refrigeration slows down metabolic activity and thus glycolysis by blood cells. (C)</p> Signup and view all the answers

Hemolysis in a blood sample can falsely elevate the results for certain analytes because:

<p>It releases intracellular components, such as enzymes, into the serum or plasma. (C)</p> Signup and view all the answers

Lipemia, or excessive fat in the blood, can interfere with laboratory assays and falsely increase hemoglobin measurements because:

<p>Lipids scatter light, interfering with spectrophotometric measurement of hemoglobin. (C)</p> Signup and view all the answers

Forcibly expelling blood from a syringe through the needle into a collection tube can cause:

<p>Hemolysis of red blood cells due to mechanical damage. (D)</p> Signup and view all the answers

Underfilling an EDTA tube can lead to red blood cell crenation because:

<p>The osmotic effect of excess EDTA draws water out of red blood cells, causing them to shrink. (B)</p> Signup and view all the answers

Expired reagents in a laboratory assay are a source of which type of error?

<p>Analytical error. (D)</p> Signup and view all the answers

What is the purpose of calibrating a laboratory instrument when using a new batch of reagents?

<p>To adjust the instrument to account for potential variations in the new reagent batch. (B)</p> Signup and view all the answers

Laboratory controls are typically run daily to:

<p>Ensure all aspects of the assay, including reagents and instrument function, are working correctly. (D)</p> Signup and view all the answers

Reporting a patient's lab results under another patient's name is an example of a:

<p>Post-analytical error. (C)</p> Signup and view all the answers

Why are reference intervals laboratory-specific?

<p>Different labs use different analyzers, reagents, and methodologies, which can affect test results. (D)</p> Signup and view all the answers

In establishing reference intervals, why are outliers typically excluded from the dataset?

<p>Outliers can skew the statistical analysis and misrepresent the healthy population's range. (C)</p> Signup and view all the answers

If a laboratory has fewer than 20 healthy animals for establishing a reference interval, what is the recommended approach?

<p>Report the lowest and highest values observed as the reference range without statistical analysis. (D)</p> Signup and view all the answers

A reference interval is statistically designed to include approximately what percentage of a healthy population?

<p>95% (C)</p> Signup and view all the answers

Diagnostic cut-offs, unlike reference intervals, are established primarily to:

<p>Differentiate between diseased and non-diseased populations, optimizing test performance for disease detection. (A)</p> Signup and view all the answers

Why is a twofold increase in serum potassium generally considered more clinically significant than a twofold increase in alanine aminotransferase (ALT)?

<p>Potassium plays a critical role in cardiac and neuromuscular function, making even moderate changes impactful. (B)</p> Signup and view all the answers

A normal creatinine level in a sick animal might be misleading, particularly in early stages of:

<p>Early kidney disease. (B)</p> Signup and view all the answers

For a disease screening test, which diagnostic property is MOST important?

<p>High sensitivity. (A)</p> Signup and view all the answers

If a test has 95% sensitivity, what is the implication regarding false negative results?

<p>The test will miss 5% of individuals who actually have the disease (false negatives). (B)</p> Signup and view all the answers

Disease prevalence in a population significantly affects which diagnostic test property?

<p>Positive and negative predictive values. (A)</p> Signup and view all the answers

Given: True Positives (TP) = 6, True Negatives (TN) = 966, False Positives (FP) = 7, False Negatives (FN) = 21. What is the accuracy of this diagnostic test?

<p>97.42% (D)</p> Signup and view all the answers

Which category of laboratory errors is most directly influenced by the blood collection process itself?

<p>Pre-analytical errors (D)</p> Signup and view all the answers

Veterinarians have the most direct control over minimizing which type of laboratory error?

<p>Pre-analytical errors during sample collection and handling (C)</p> Signup and view all the answers

Using a syringe and needle for blood collection, compared to vacutainers, introduces a slightly increased risk of which pre-analytical error?

<p>Increased time between collection and anticoagulation (C)</p> Signup and view all the answers

The presence of clotting factors is a key distinction when comparing:

<p>Plasma and serum (A)</p> Signup and view all the answers

What is the primary role of the gel within a serum separator tube (SST)?

<p>To create a physical barrier between serum and blood cells after centrifugation (A)</p> Signup and view all the answers

Why is it crucial to allow complete clot formation before centrifuging a red top tube?

<p>To ensure clotting factors are fully consumed and serum is properly separated (A)</p> Signup and view all the answers

For which of the following analytes is a red top tube generally considered MOST appropriate for collection?

<p>Serum Biochemistry profile (e.g., electrolytes, liver enzymes) (B)</p> Signup and view all the answers

Serum separator tubes (SSTs) are generally discouraged for therapeutic drug monitoring because:

<p>The gel can interfere with drug assay results. (A)</p> Signup and view all the answers

Ethylene diamine tetraacetic acid (EDTA) prevents blood coagulation by which mechanism?

<p>Chelating calcium ions (D)</p> Signup and view all the answers

A critically low calcium result from a purple top (EDTA) tube should raise suspicion of:

<p>Sample contamination with EDTA, artificially lowering calcium (D)</p> Signup and view all the answers

Why are purple top (EDTA) tubes considered the preferred choice for performing cell counts?

<p>EDTA preserves cell morphology and prevents cell clumping. (A)</p> Signup and view all the answers

Lithium heparin is often favored over sodium heparin for general chemistry profiles primarily because:

<p>Sodium heparin can falsely elevate sodium measurements. (A)</p> Signup and view all the answers

For 'stat' (immediate) chemistry testing, lithium heparin tubes offer an advantage because:

<p>Plasma can be analyzed directly without waiting for clot formation. (C)</p> Signup and view all the answers

In situations with very limited blood volume, a green top (heparin) tube is versatile because it allows for:

<p>Sequential CBC and chemistry testing from a single small sample. (B)</p> Signup and view all the answers

Sodium citrate (blue top tubes) is specifically used for coagulation testing because it:

<p>Provides a strong anticoagulation that can be reversed by adding calcium. (A)</p> Signup and view all the answers

If a glucose sample in a plain tube is not promptly processed at room temperature, glucose levels are expected to:

<p>Decrease due to cellular glycolysis. (A)</p> Signup and view all the answers

Refrigeration helps preserve glucose levels in blood samples by:

<p>Inhibiting enzymatic activity, including glycolysis. (C)</p> Signup and view all the answers

Hemolysis falsely elevates results for certain analytes because:

<p>It releases intracellular components into the plasma or serum. (C)</p> Signup and view all the answers

Lipemia can falsely increase hemoglobin measurements in certain assays due to:

<p>Light scattering interference in spectrophotometric assays. (D)</p> Signup and view all the answers

Using expired reagents in a laboratory assay is a source of which type of error?

<p>Analytical error (C)</p> Signup and view all the answers

The primary purpose of calibrating a laboratory instrument with a new batch of reagents is to:

<p>Ensure the instrument provides accurate results with the new reagent lot. (A)</p> Signup and view all the answers

Reporting a patient's laboratory results under a different patient's name is an example of a:

<p>Post-analytical error. (C)</p> Signup and view all the answers

Reference intervals are laboratory-specific primarily because:

<p>Different laboratories use varied analyzers, reagents, and methodologies. (C)</p> Signup and view all the answers

In establishing reference intervals, outliers are typically excluded from the dataset because they:

<p>Can skew the statistical analysis and misrepresent the healthy population. (D)</p> Signup and view all the answers

If a laboratory has fewer than 20 healthy animals available for establishing a reference interval, a recommended alternative approach is to:

<p>Use non-parametric statistics on the limited data, reporting observed ranges. (D)</p> Signup and view all the answers

A reference interval is statistically designed to encompass approximately what percentage of a healthy population?

<p>95% (A)</p> Signup and view all the answers

Diagnostic cut-offs, in contrast to reference intervals, are primarily established to:

<p>Discriminate between diseased and non-diseased populations. (A)</p> Signup and view all the answers

A normal creatinine level in a sick animal can be misleading, particularly in the early stages of:

<p>Kidney disease. (C)</p> Signup and view all the answers

For a disease screening test, which diagnostic property is MOST important to maximize?

<p>Sensitivity (C)</p> Signup and view all the answers

If a diagnostic test has a 95% sensitivity, this implies that:

<p>5% of individuals with the disease will test negative (false negatives). (D)</p> Signup and view all the answers

Given: True Positives (TP) = 6, True Negatives (TN) = 966, False Positives (FP) = 7, False Negatives (FN) = 21. Calculate the accuracy of this diagnostic test.

<p>97.7% (D)</p> Signup and view all the answers

Which of the following scenarios represents a post-analytical error?

<p>Entering laboratory results into the wrong patient's electronic record. (C)</p> Signup and view all the answers

An ELISA test for a specific disease can be adjusted to increase sensitivity. What is the MOST likely consequence of increasing the sensitivity of this test?

<p>An increase in false positives and a decrease in false negatives. (D)</p> Signup and view all the answers

Consider a scenario where a veterinarian is screening for a rare disease in a general pet population. Which test characteristic is MOST crucial for minimizing missed cases of this rare disease?

<p>High Sensitivity (A)</p> Signup and view all the answers

In the context of diagnostic testing, what does 'bad data is worse than no data' fundamentally imply?

<p>Inaccurate laboratory results can lead to incorrect diagnoses and harmful treatments. (D)</p> Signup and view all the answers

For which patient condition are chemistry results MOST likely to be falsely elevated due to hemoconcentration?

<p>Dehydration (B)</p> Signup and view all the answers

Enzyme activities in a blood sample, measured in U/L, are MOST susceptible to changes due to:

<p>Sample storage temperature and time. (B)</p> Signup and view all the answers

Reference intervals for laboratory tests are ideally established using samples from:

<p>A large cohort of clinically healthy animals from the target population. (A)</p> Signup and view all the answers

The primary reason each laboratory should ideally establish its own reference intervals is:

<p>To account for variations in analytical methods and patient populations. (B)</p> Signup and view all the answers

In a box and whisker plot of platelet counts, outliers are identified and should be investigated primarily because they:

<p>May indicate errors in measurement or represent pathological conditions. (D)</p> Signup and view all the answers

When establishing reference intervals, if a laboratory observes platelet clumps on a blood smear associated with unusually low platelet counts, these data points should be:

<p>Excluded from the reference interval calculation as pre-analytical errors. (D)</p> Signup and view all the answers

Which type of laboratory error is LEAST likely to be directly controlled by a veterinarian if samples are sent to a reference lab?

<p>Analytical errors occurring during sample processing. (C)</p> Signup and view all the answers

What can occur if a blood sample is collected too slowly using a small-gauge needle?

<p>Activation of platelets and potential clot formation. (A)</p> Signup and view all the answers

What is the primary advantage of using vacutainers directly for blood collection, as practiced in human medicine, compared to using a needle and syringe?

<p>Direct transfer of blood into the collection tube, minimizing pre-analytical errors. (D)</p> Signup and view all the answers

Serum differs from plasma primarily because serum lacks:

<p>Clotting factors, which are consumed during clot formation. (A)</p> Signup and view all the answers

What role does the gel separator play in serum separator tubes (SSTs)?

<p>Creates a barrier between serum and blood cells after centrifugation. (D)</p> Signup and view all the answers

What is the most critical step to take before centrifuging a red top tube after blood collection, and why?

<p>Allowing the blood to clot completely to ensure proper serum separation. (C)</p> Signup and view all the answers

For which analyte is a red top tube generally MOST appropriate?

<p>Therapeutic drug monitoring. (C)</p> Signup and view all the answers

Why are serum separator tubes (SSTs) generally NOT recommended for therapeutic drug monitoring?

<p>The gel in SSTs can absorb certain drugs, leading to falsely decreased measurements. (D)</p> Signup and view all the answers

What is the mechanism by which EDTA prevents blood coagulation?

<p>Chelating calcium, which is essential for the coagulation cascade. (B)</p> Signup and view all the answers

A chemistry profile from a purple top (EDTA) tube shows a critically low calcium level (e.g., 2 mg/dL). What is the MOST likely explanation?

<p>The EDTA in the tube chelated the calcium, leading to a falsely low result. (B)</p> Signup and view all the answers

Why is a purple top (EDTA) tube considered a good choice for performing cell counts?

<p>EDTA preserves cell morphology by preventing clot formation. (C)</p> Signup and view all the answers

Lithium heparin is often preferred over sodium heparin for general chemistry profiles primarily because:

<p>Sodium heparin can falsely elevate sodium levels in the sample. (B)</p> Signup and view all the answers

How does refrigeration help preserve glucose levels in blood samples?

<p>By slowing down the metabolic activity of blood cells. (C)</p> Signup and view all the answers

A laboratory is evaluating a new diagnostic test for a rare disease. Which method would be MOST suitable for comparing the results of the new test with those of an established, highly accurate test when evaluating its performance?

<p>Measuring the sensitivity and specificity of the new test relative to the established test. (D)</p> Signup and view all the answers

Which measure is MOST critical to evaluate when choosing a screening test designed to identify a disease in its early stages, especially when the disease has severe consequences if left untreated?

<p>Maximizing the test's sensitivity to identify as many true positives as possible and minimize false negatives. (A)</p> Signup and view all the answers

A veterinary clinic is using a point-of-care instrument to measure blood glucose. The instrument has a built-in quality control system that performs regular internal checks. Despite this, what additional step is MOST important to consistently ensure the reliability and accuracy of the instrument's measurements?

<p>Conducting weekly external quality control checks using control solutions with known values. (A)</p> Signup and view all the answers

Flashcards

What is serum?

Liquid remaining after blood has been allowed to clot.

What are preanalytical errors?

Errors that occur before the sample is analyzed, such as incorrect collection or handling.

What are analytical errors?

Errors that occur during the actual testing or analysis of the sample.

What are postanalytical errors?

Errors that occur after the sample has been analyzed, such as transcription mistakes.

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Why place the needle cleanly in vein?

To avoid clotting and hemolysis.

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Why avoid prolonged draw time?

Platelets activate and clump, affecting test results.

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What is the benefit of using vacuutainer system?

Blood goes directly into the tube, minimizing exposure to air and contamination.

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What is special about purple top tubes?

Purple top tubes contain EDTA, which strongly chelates calcium and are used for cell counts.

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What is special about green top tubes?

Green top tubes contain heparin, which activates antithrombin and are used for certain cell counts or chemistries.

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What is special about blue top tubes?

Blue top tubes contain citrate that weakly chelates calcium and are used for coagulation tests.

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What is special about gray top tubes?

Gray top tubes contain potassium oxalate and sodium fluoride used for glucose determinations.

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How does hemolysis affect lab data?

Hemolysis releases substances like AST and LDH from red blood cells, interfering with test results.

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How does lipemia affect lab data?

Lipemia causes turbidity, which interferes with hemoglobin measurements.

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Why avoid expired reagents?

Expired reagents can lead to inaccurate test results.

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What is the purpose of calibrators?

Instruments need calibration with known values to produce accurate results.

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What is the purpose of controls?

Test the consistency and reproducibility.

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What impact reference Intervals?

Values depend on methodology, reagents, and temperature.

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What affects reference values?

Age and breed.

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What is the issue with minor changes in lab data?

It may not be significant

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What is sensitivity?

Percentage of true positives.

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What is specificity?

Percentage of true negative results.

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What is the Efficiency?

Percentage of patients correctly classified by the test.

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What is PPV?

Probability that a positive test is actually positive.

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What is NPV?

Probability that a negative test is actually negative.

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What happens when cut-off is increased?

Sensitivity decreases, specificity increases.

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What happens when cut-off is lowed?

Sensitivity increases, specificity decreases.

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Which is a better screening test?

A highly sensitive test.

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What's a good confirmatory tests?

A highly specific test.

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Collection Errors

Things can go wrong when you're collecting a sample, such as poor technique or a fractious animal.

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Benefits of Larger Veins

Larger veins provide faster flow, reducing the likelihood of platelet aggregation and clot formation during collection.

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Risks of Small Veins

Small veins can collapse under negative pressure, leading to red cell breakage (hemolysis) and platelet activation.

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Serum

The liquid portion of blood after it has clotted.

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Plasma

Liquid portion of blood when clotting is prevented by an anticoagulant.

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Serum Separator Tubes (SST)

These tubes have a gel that separates serum from the clot after centrifugation.

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Timing of Centrifugation

Allow full clot formation before centrifugation to avoid a spongy clot that traps serum.

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Substances Measurable in Serum

Glucose, sodium, potassium, albumin, cholesterol, and triglycerides.

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Preferred Tubes for Hormone/Drug Assays

Plain red top tubes or white top tubes are used. SST tubes are avoided due to potential interference from the gel or clot activator.

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Blood Sample Settling

If a blood sample settles out due to gravity it can be remixed.

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High Potassium

A high potassium level in a sample indicates it was contaminated with EDTA.

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Heparin Mechanism

Heparin activates antithrombin, a natural anticoagulant, preventing clot formation through a different mechanism than calcium chelation.

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Type of Heparin

Lithium heparin is preferred over sodium heparin to avoid falsely elevated sodium levels in chemistry profiles.

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Benefits of Heparin Use

Heparinized samples don't need to clot, good for urgent tests.

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Versatility of Green Top Tubes

You can perform both cell counts and chemistry on a single small volume sample.

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Glycolysis

Glycolysis, or the consumption of glucose by cells in the sample.

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Function of Sodium Fluoride

It will prevent glycolysis, useful when precise glucose measurement is needed.

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Glucose Consumption Rate

Glucose decreases by 10% per hour at room temperature.

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Enzymes Released by Hemolysis

AST, LDH

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Poorly Labeled Samples

The sample may have the wrong animal name, or lack sufficient information.

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Damage During Transport

Freezing or overheating can cause red cell lysis or sample degradation.

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Proper Filling Technique

Blood should be drawn into the tube via vacuum, rather than forced, helps avoid lysing cells.

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Effects of Underfilling EDTA Tubes

EDTA is osmotic and can cause red cells to shrink if the tube is underfilled.

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Cross-Contamination Example

Contamination of a red top tube with EDTA from a previous sample.

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When to Calibrate

Instrument needs recalibration when you use a new lot of reagent.

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Analyzer Linearity

A range above or below which results may not be accurately measured by an analyzer.

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Consequences of Post-Analytical Errors

Transcription errors, mislabeled samples, or incorrect data entry can lead to medical mistakes.

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Hemo Concentration

A high albumin or sodium level because the animal is dehydrated.

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Reference Intervals

Ranges of values obtained from a population of healthy animals.

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Challenges of Reference Intervals

Hard to collect, not always healthy.

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Data distribution

If smaller, use specific statistics

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Using data for interpretations.

Look at reference intervals.

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Diagnostic Criteria

Guidelines to look at values to suggest disease that may or may not be normal.

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High Alkaline Phosphatase in young patients.

Bones are remodeling.

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Breed Specific Differences

There are lower cell counts in canines.

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Study Notes

Factors Affecting Interpretation of Lab Data

  • This topic focuses on the various elements influencing the accuracy and reliability of laboratory data interpretation.

Learning Objectives

  • Comprehend the color-coding for blood collection tubes.
  • Define serum and plasma and know the distinctions.
  • Identify the appropriate collection tubes for mammalian CBC, coagulation assays, serology tests, and chemistry profiles.
  • Recognize the importance of filling collection tubes in the correct order, specifically avoiding the filling of an EDTA tube before a red top tube.
  • Identify pre-analytic, analytic, and post-analytic sources of error.
  • Explain problems stemming from the over or under-filling of collection tubes.
  • Know how to derive and use measures such as sensitivity, specificity, PPV, NPV, and accuracy.

Sources of Error

  • Lab data interpretation is subject to preanalytical, analytical, and postanalytical errors.
  • As a veterinarian, you have the most control over pre-analytical errors.
  • Errors can occur during collection, shipping, etc.
  • Analytical errors can occur as the sample is being run.
  • Post-analytical errors occur during the reporting of results.

Preanalytical Errors

  • Poor collection technique can cause preanalytical errors.
  • Clotting or hemolysis is avoided by placing needles directly into the vein.
  • Blood collection is easier and more effective when using large veins rather than small veins.
    • Large veins are less prone to collapse.
    • Large veins allow for faster blood flow.
  • Platelet activation and clumping are avoided by limiting draw time.
  • The Vacutainer system can be used for blood collection.
    • It allows the blood to go straight into the tube.
  • The use of incorrect tube types can create error.
    • Examples: incorrect or no anticoagulant, blood allowed to clot
  • Other preanalytical errors include:
    • Interfering substances such as hemolysis.
      • AST and LDH can be found in red blood cells.
    • Lipemia.
      • Turbidity increases hemoglobin.
    • Improperly labeled samples, damage during transport.
  • Errors can be introduced when filling tubes.
    • The sample should not be forced through the needle; instead, let the tube fill via vacuum.
    • Incorrectly filled tubes can create errors.
      • Anticoagulant dilution, excess EDTA can have osmotic effects
    • Cross-contamination is a potential error.
      • Remove the cap from the tube and fill directly from the syringe to avoid this.
  • Struggling with the animal during collection can affect the sample.
  • Jugular veins give cleaner samples because of faster flow but the animal may not allow collection from this site.
  • Small veins can collapse, which can break red cells and activate platelets, leading to hemolysis.
  • In the human world blood is taken directly from the vein into the tube, but in veterinary medicine there is a step in between the vein and the tube.
  • You have to know which tube you need for your sample.
  • If you have no anticoagulant, then the blood will clot, because an anticoagulant will prevent clotting.

Vacutainer Tubes With Anticoagulants

  • Tubes with anticoagulants include purple, green, and blue tops.
  • Purple top tubes contain EDTA.
    • EDTA strongly chelates calcium.
    • Purple tops can be used for cell counts in blood or fluid and plasma hormone and ammonia testing.
    • Contamination with EDTA can cause falsely low calcium and high potassium levels.
  • Green top tubes contain heparin.
    • Heparin activates antithrombin.
    • Green tops are used for cell counts, especially in some reptiles, and plasma hormone and stat chemistry testing.
    • Can be lithium heparin or sodium heparin and lithium heparin is best if you're going to do any kind of chemistry testing.
    • The sample does not have to wait to clot with lithium heparin.
    • It can be used for both chemistries and cell counts.
  • Blue top tubes contain citrate.
    • Citrate more weakly chelates calcium.
    • Citrate tubes are used for plasma coagulation tests.
    • Calcium can be added back into sodium citrate and coagulation tests can work.

Gray Top Tubes

  • Gray top tubes contain potassium oxalate as an anticoagulant and sodium fluoride.
  • The tubes inhibit glycolysis and are used for glucose determinations.
  • The red cells and white cells will eat any glucose, so glucose levels drop if samples sit around.
  • These tubes prevent the red cells from eating the glucose if a precise value is needed.
  • Glucose gets consumed at room temperature at about 10% per hour.
  • Little consumption happens when refrigerated and glucose survives pretty well after being spun down.

Analytical Sources of Error

  • Analytical errors can be caused by expired reagents.
  • Failure to calibrate the assay as needed can cause analytical error.
    • Whenever the reagent lot is changed, the instrument needs to be calibrated.
  • Analyzer limitation can create analytical error.
    • If the concentration of an analyte exceeds the accuracy of the assay, it can cause analyzer error.
  • Another analytic error can be created when the method of testing is not appropriate for the species.
  • When you get a new lot of reagent, you need to calibrate the instrument using a calibrator with a known result to tell the instrument how to run.
  • You have to know your analyzer linearity and how high it can go.

Calibrators and Controls

  • These tools ensure reliability and consistency of assay results.
  • Calibrators configure an assay using a sample with a known value.
    • The instrument is "taught" to produce accurate results.
    • This is done intermittently.
  • Controls verify the recovery level of standardized reagents.
    • Controls evaluate an assay's consistency or reproducibility.
    • Controls are generally done daily.

Post-Analytical Sources of Error

  • Transcription errors and results reported to the wrong patient can lead to post-analytical errors.
  • It is better to have no data than bad data.
    • The lab should be contacted if the data does not make sense.
  • Errors from mislabeled samples can go into the medical record of the wrong animal.

Units of Measure

  • Percentages and concentrations are used.
    • Chemistry results are higher in concentrated plasma.
      • Dehydration will increase albumin, sodium, and chloride concentrations, along with the PCV (% of RBCs in blood).
    • Percentage of lymphocytes in a CBC.
      • Increased lymphs VS decreased neutrophils?
  • Enzyme activities in U/L are affected by both the amount of enzyme and the sample age.

Reference Intervals

  • Reference intervals define a healthy population.
  • Values depend on the methodology.
    • Different reagents, cofactors, substrates, temperature, etc.
  • Each lab should have its own reference intervals.
    • Intervals are ideally based on the values from approximately 120 healthy patients.
  • Reference intervals are specific for the lab.
  • It is hard to come up with 120 healthy animals.
  • The data needs to have a normal distribution or a skewed distribution and outliers need to be identified.

Establish Rejection Criteria

  • Box and Whisker plots can reveal outliers.
    • For example, a K9 platelet count with outliers caused by platelet clumps.
  • Outliers and data in samples that have interfering factors should be excluded.

Reference Interval Statistics

  • Statistics will vary depending on factors like the number of healthy animals and whether the data is normally distributed.
  • No reference intervals are available if <20.
  • Present mean value and a histogram of observed data in these scenarios.
  • Reference intervals reflect approximately 95% of the population.
  • The statistics you choose will depend on whether you have a normal Gaussian distribution, a nice bell curve, in which case you can use mean and two standard deviations.
  • If you have less than 40 patients, then you may have to use different, what they call non-parametric statistics.

Implications

  • The reference interval reflects ~95% of the population.
    • 2.5-5% of normal patients will be outside this interval.
  • Those reference intervals are going to be based on healthy animals.

Diagnostic Cut-Off

  • This is an alternative to the reference interval.
  • It sets an assay value as likely to be diagnostic for the disease of interest.
  • It looks at animals with the disease instead of healthy animals.
  • Diagnostic cutoffs look at animals that have a disease versus other animals that may or may not be healthy.
  • Recommendations are given on what value you should strongly suspect that you have the disease you're looking for.

Reference Intervals

  • Age and breed affect reference values.
    • Increased alk phos, Ca, & phos is one such example in growing animals.
    • Lower WBC and platelet counts in greyhounds are another.
  • Minor changes may not be significant.
    • A 2x increase in K is dangerous.
    • A 2x increase in ALT may be less dangerous.
  • Sick animals may have normal test results.
    • Normal creatinine can be seen with early renal disease.
  • Young animals will have high alk phos, high calcium, high phosphorus because of just bone remodeling.
  • Select for one year to six years when doing reference intervals to try to avoid some of these outliers.
  • You have to get a feel for when is a change significant as you look at more data.

Assay Diagnostic Properties

  • Important properties include:
    • Sensitivity
    • Specificity
    • Positive Predictive Value
    • Negative Predictive Value

Sensitivity

  • Sensitivity is the percent of true positives.
    • True positives / all patients with a specified disease.
    • If an assay is 95% sensitive, it has 95% true positives and a 5% false negative rate.

Specificity

  • Specificity is the percent of true negative results.
    • True negatives / all patients without a specified disease.
    • If an assay is 95% specific, it has 95% true negatives and a 5% false positive rate.

Efficiency (Accuracy)

  • Represents the percentage of patients that are correctly classified by the test.
    • Calculated as True positives + True negatives / total patients examined

Predictive Values

  • These are affected by sensitivity, specificity, and disease prevalence.
  • PPV: Probability that a positive test is correct.
    • Calculated as true positives / all positives
  • NPV: Probability that a negative test is correct.
    • Calculated as true negatives / all negatives
  • As a veterinarian, what you really want to know are your predictive values.

Diagnostic Criteria

  • Changing the diagnostic criterion will effect interpretation.
    • A 100% sensitivity, but lower specificity could mean a poor PPV because there are many FP results.
    • A 100% specificity but lower sensitivity could mean a poor NPV because there are many FN results.
  • A test with high specificity is a good confirmatory test and will increase the PPV.
    • A positive result in this case means the animal likely has the disease in question.
    • NPV in this case is worse
    • A negative result does not rule out disease.
  • Sensitivity is a measure of true positives.
  • Specificity is going to be the true negatives.
  • Accuracy is of all my population those with disease those without how many did we get right.
  • Diagnostic criteria/ cutoffs can be tweaked.
  • Tweaking can change the sensitivity or specificity.
  • When a company is making a test, they can tweak where they put these cutoffs.
  • If you increase sensitivity, you give up specificity.
  • If you have a high sensitivity, then you're going to have a poor positive predictive value.
  • If you have a highly specific test you're going to have a poor negative predictive value because you have a lot of false negatives.
  • A screening test should have a higher sensitivity.
  • Specific tests make good confirmatory tests, negatives not as reliable.
  • If you get a negative on a highly sensitive test, you can take that to the bank.
  • A highly sensitive test is good for screening because you cast a broad net.

Serum vs Plasma

  • If the sample clots, the liquid that remains is called serum.
  • If you put your blood into an anticoagulant and it doesn't clot, then the liquid that remains is plasma.
  • The part of the difference between serum and plasma is clotting factors.
  • Proteins that comprise our clotting factors are going to become incorporated into the clot.
  • Protein concentration will be slightly lower in serum than plasma.

Tube Types

  • There is some difference between the brands of tubes.
  • Red top tubes are used, but sometimes tiger top tubes are used.
  • The SST tube has a gel in it that is sitting on the bottom of the tube, and then you would put the blood on top.
  • When you centrifuge this sample, the gel will migrate up, and it inserts itself between the clot and the serum.
  • It's important to let it fully clot before you spin it or the material will be like a sponge.
  • You can run a chemistry test like glucose on this tube.
  • You can put things like urine samples in these sterile tubes.
  • White top tubes are an alternative to red top tubes.
  • Plain red top tubes or white top tubes are better for running some drug assays.

Hemolysis

  • This can be due to red cell breakage.
  • As you break those red cells, things that should be in their cytoplasm are going to be in your sample.
  • The red cells tell you if there is hemoglobin there and you have to sort out whether this happened while you were collecting the sample or is it happening in the animal.
  • The animal will be anemic if it is happening in the animal.

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