MICI: Molecular Techniques

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Questions and Answers

Which step of the PCR process involves the binding of primers to target DNA?

Annealing (correct)

Amplification

Denaturation

Extension

What is a key advantage of using 16S rRNA gene sequencing for bacterial identification?

It can identify slow-growing or non-viable organisms. (correct)

It requires culturing of bacteria.

It cannot differentiate between closely related species.

It analyzes only viable organisms for accuracy.

What method is primarily used in outbreaks to genotype bacteria like E.coli O157?

Pulse Field Gel Electrophoresis (correct)

Whole Genome Sequencing

Real-time PCR

Nucleic Acid Extraction

What is the primary purpose of nucleic acid extraction and purification in molecular diagnostics?

To prepare samples for subsequent testing Signup and view all the answers

Which nucleic acid amplification technique can quantify nucleic acids during the amplification process?

Real-time PCR Signup and view all the answers

Which statement accurately describes the regions targeted in 16S rRNA gene sequencing?

Both conserved and divergent regions are utilized. Signup and view all the answers

What is the primary reason PCR requires specifically designed primers?

To ensure high specificity for the target organism. Signup and view all the answers

What makes the 16S rRNA gene a reliable target for bacterial identification?

It is essential for protein synthesis in all bacteria and evolves slowly. Signup and view all the answers

Why are nucleic acid amplification tests (NAAT), like PCR, beneficial in clinical diagnostics?

They can detect both live and dead organisms. Signup and view all the answers

For which primary purpose is pulsed field gel electrophoresis (PFGE) utilized?

For typing and comparing bacterial strains in outbreak investigations. Signup and view all the answers

What advantage does whole genome sequencing (WGS) provide in outbreak investigations?

It allows for the mapping of organism transmission networks. Signup and view all the answers

Which of the following typing systems is based solely on genetic analysis rather than metabolic or biological activities?

Pulsed field gel electrophoresis. Signup and view all the answers

What is a primary advantage of using PCR in clinical diagnostics?

It can detect very small quantities of pathogen DNA/RNA. Signup and view all the answers

Which of the following statements best describes the role of 16S rDNA sequencing in clinical diagnostics?

It is essential for identifying slow-growing or non-culturable bacteria. Signup and view all the answers

In what scenario is PCR especially critical according to the information provided?

When immediate diagnosis and treatment are essential. Signup and view all the answers

How does 16S rDNA sequencing contribute specifically to the treatment of UTIs?

It identifies the specific bacterial strain involved. Signup and view all the answers

What is an essential application of PCR during outbreak investigations?

Detecting viral RNA to confirm diagnoses. Signup and view all the answers

Which of the following pathogens can be detected with 16S rDNA sequencing?

Bacteria like Chlamydia trachomatis. Signup and view all the answers

What is a major limitation of traditional culturing methods compared to molecular techniques?

They can only identify pathogens that can be easily cultured. Signup and view all the answers

Which of the following describes a key benefit of rapid PCR testing in a clinical setting?

It significantly reduces the time to diagnosis. Signup and view all the answers

What is an important distinction between viruses and bacteria regarding stability?

Viruses are generally more stable than bacteria under laboratory conditions. Signup and view all the answers

Which component is crucial for the specificity of PCR in identifying pathogens?

The primers designed to target specific sequences. Signup and view all the answers

Study Notes

Applications of Molecular Techniques in Clinical Diagnostics

Molecular techniques like PCR and 16S rDNA sequencing are crucial for rapid and accurate identification of pathogens
PCR is used for outbreak investigations, diagnostic accuracy, and detecting viral and bacterial infections
- PCR can detect pathogens during disease outbreaks (e.g., Ebola, COVID-19)
- PCR is used to detect viral load in HIV and track viral progression
- PCR can detect viruses like Influenza, RSV, and measles, which are mentioned in the test review sheet
Advantages of PCR:
- Sensitivity: can detect small amounts of a pathogen's DNA/RNA
- Speed: PCR provides results within hours, while traditional culture methods can take days
16S rDNA sequencing is vital for bacterial identification, particularly when culturing is difficult or slow.
- 16S rDNA sequencing can rapidly identify specific bacterial strains, guiding more targeted antibiotic treatment
- 16S rDNA sequencing can detect bacteria that are difficult or impossible to grow in culture
16S rDNA sequencing advantages:
- Detects Slow or Non-Growing Organisms Useful in detecting pathogens that don't grow easily in cultures (e.g., Mycobacterium)
- No Need for Viable Organisms 16S rDNA PCR can work on dead bacterial cells
Pulse Field Gel Electrophoresis (PFGE) was used for comparing bacterial strains during outbreak investigations but is being replaced by Whole Genome Sequencing (WGS)
- PFGE played a key role in investigating foodborne outbreaks (e.g., E.coli O157)
- WGS provides more detailed genetic information and is now often used to track outbreaks
Real-Time PCR quantifies DNA in real-time as it amplifies, making it useful for viral load monitoring and bacterial identification
- Real-time PCR is used to monitor viral load in HIV and assess the effectiveness of antiretroviral therapy
- Real-time PCR can detect and quantify viral particles in infections like rotavirus and norovirus
Molecular techniques have profound implications for diagnosing infections:
- Reduce Diagnosis Time: Faster than traditional culturing methods
- Increase Sensitivity and Specificity: Can detect latent infections and differentiate between viral and bacterial pathogens
- Enable Detection of Multiple Pathogens Simultaneously: Helpful for diagnosing co-infections.
Molecular techniques play a pivotal role in modern diagnostics:
- Optimize Treatments: by targeting the specific pathogen causing the infection
- Prevent the Spread of Infectious Diseases: through timely intervention, particularly during outbreaks

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Description

Explore the applications of molecular techniques in clinical diagnostics, focusing on PCR and 16S rDNA sequencing. Learn how these methods enhance pathogen identification and improve diagnostic accuracy, especially during outbreaks. Understand the advantages of PCR in detecting various viral and bacterial infections.