MICR 111 Specimen Collection, Culture and Inoculation Technique PDF

MICR_111 NATHANIEL VERON M. CASAMINA, MSMT, RMT, ASCP | 3MT03 3RD YEAR, 1ST SEMESTER BY: MANALO, A. & MILAR, K. [TRANS] LESSON #4: SP...

MICR_111 NATHANIEL VERON M. CASAMINA, MSMT, RMT, ASCP | 3MT03 3RD YEAR, 1ST SEMESTER BY: MANALO, A. & MILAR, K. [TRANS] LESSON #4: SPECIMEN COLLECTION, CULTURE AND INOCULATION TECHNIQUE OUTLINE Hematological Analysis (3rd tube) I. Specimen Collection Cryptococcus II. Transport Media neoformans - Fungi III. Blood Culture Collection that causes meningitis. A. Transfer Blood to Culture Media India ink is used to stain IV. Common Specimens in Microbiology them. V. Take Note VI. Specimen Labeling VII. Specimen Transportation Collection of CSF - VIII. Preservatives and Anticoagulant Spinal Tap IX. Specimen Prioritization Ear: Inner Clean ear canal with Sterile, screw-cap X. Specimen Rejection mild soap, aspirate tube or anaerobic XI. Sputum Criteria fluid with transport system needie if eardrum intact; use Swab if SPECIMEN COLLECTION eardrum ruptured Specimens for microbiology cultures should be collected in Ear: Outer Remove debris of Swab transport sterile containers except for stool specimens. crust from ear canal system o Collected in sterile containers inorder to with saline-moistened swab; rotate swab in avoid cross contamination. outer canal Swab Eye: Conjunctiva Sample both eyes; Swab transport o Submit 2 samples use separate swabs system o Smear (1st sample) and Culture (2nd moistened with sterile sample) saline Tips are modified already and before cotton tips are used Eye: Corneal Instill local anesthetic, Agar available at but they found out that cottons are toxic to some scrapings scrape with sterile bedside microorganisms because it contains fatty acids that kills spatula and inoculate the microorganisms. directly to agar Alternative to Cotton Tips with: Dacron, rayon, or Feces Collect directly into Clean, leakproof container, avoid container or enteric calcium alginate (SARS-CoV-2) contamination with transport system urine Table No. 1 Specimen Collection Guidelines Fungal scrapings Wipe nails or skin Clean, screw-cap SPECIMEN COLLECTION GUIDELINES Hair/nail/skin with alcohol container SPECIMEN PATIENT CONTAINER/ PREPARATION MINIMUM Hair: 10-12 hairs with QUANTITY shaft intact Blood Culture Disinfect skin with Blood culture media alcohol and iodine or set aerobic and Nails: Clip affected chlorhexidine anaerobic bottles) or area Vacutainer tube with SPS/adults 20 ml. per Skin: Scrape skin at set; children 5-10 mL outer edge of lesion per set Genitalia: Remove mucus Swab transport Body fluids Disinfect skin before Sterile, screw-cap Cervix/vagina before collection: do system or JEMBEC (abdominal, amniotic, needle aspiration tube or anaerobic not use lubricant on transport ascites, bile, joint, transport system/21 speculum; swab System pericardial, pleural) ml. endocervical canal or Catheter tips, IV (Foley Disinfect skin before Sterile, screw-cap vaginal mucosa. catheters not cultured) removal container Genitalia: Flexible swab Swab transport Cerebrospinal Fluid Disinfect skin before Sterile, screw-cap Urethra inserted 2-4 cm into system or JEMBEC aspiration tube/bacteria 21 mL, urethra forese transport system Collected in 3 tubes: fungi 22 mL, AFB 22 or collect discharge mL, virus 21 mL. Lesion/wound/ Wipe area with sterile Immunology and Clinical abscess saline or alcohol Chemistry (1st tube) Lesion/wound/ Swab along outer Swab transport abscess: Superficial edge system Microbiology Culture Lesion/wound/ Aspirate with needle Anaerobic transport and Sensitivity abscess: Deep and syringe system Microbiology (2nd tube) 1 TRINITY UNIVERSITY OF ASIA – COLLEGE OF MEDICAL TECHNOLOGY | 3MT03  LESSON #4: SPECIMEN COLLECTION, CULTURE AND INOCULATION TECHNIQUE Respiratory tract: Rinse mouth or Sterile, screw-cap BLOOD CULTURE COLLECTION lower bronchial gargle with water, container Aseptic technique (30 seconds) specimens: Instruct to cough o 70% alcohol followed by 2% tincture of Sputum deeply into container iodine Respiratory tract: Insert premoistened Swab transport o If allergic to Iodine you can use : upper: swab with sterile system Nasal saline 1 inch into Chlorhexidine gluconate and nares Benzalkonium chloride Respiratory tract: Insert flexible swab Swab transport Blood Volume upper: through nose into system or direct o Pediatric sample 5-10mL per set Nasopharynx posterior inoculation to media o Adult 20mL per set nasopharynx, rotate Skin contaminants: for 5 s o CoNS (Coagulase-negative staphylococc)i, Respiratory tract: Swab posterior Swab transport Micrococcus and Propionibacterium upper: Throat pharynx, tonsils, and system Need to follow proper aseptic technique = to avoid cross Inflamed area contamination which leads to false diagnosis. Tissue Disinfect skin; do not Anaerobic transport allow tissue to dry system or sterile Blood Culture Bottle Color Top = Anaerobic (Violet), out; it screw-cap container Aerobic (Yellow), Pediatric (Pink) necessary, moisten Blood Culture : Yellow Top (1st collected) = with with sterile saline Sodium Polyanethole Sulfonate (SPS) Urine: Clean-catch Clean external Sterile, screw-cap ○ Gold top can’t be used because it is a serum midstream genitalia; begin container of urine and it does not have an anticoagulant. voiding; after several transport kit/2-3 mL mL have passed, collect midstream without stopping flow of urine Catheter Clean urethral area, Sterile, screw-cap insert catheter, and container of urine allow first 15 ml to transport kit pass; collect remainder Indwelling catheter Disinfect catheter Sterile, screw-cap collection port, container of urine aspirate 5-10 mL. transport kit with needle and syringe Suprapubic aspirate Disinfect skin, Stenie, screw-cap aspirate with needle container of and syringe anaerobic transport through abdominal system wall into full bladder TRANSFER BLOOD TO CULTURE MEDIA Once you collected the blood sample from your culture bottle into your syringe. Remove the needle and drop by drop the blood into the culture media. Or you can use an adapter for blood culture bottle so you can drop the blood like a soy sauce. TRANSPORT MEDIA It provides a controlled environment to maintain the viability of the organism during transport. Example of transport medium: o Cary blair medium o Amies medium COMMON SPECIMENS IN MICROBIOLOGY o Stuart medium Urine (UTI) o Transgrow o Clean catch midstream o JEMBEC o Cleansing of genitalia is required prior to Sometimes a charcoal is collection. added, because microorganism require different ▪ Cleansing of genitalia is important environment. especially if collecting urine from Charcoal - added to Neisseria gonorrhoeae and foley catheter. Bordetella pertussis. 2 TRINITY UNIVERSITY OF ASIA – COLLEGE OF MEDICAL TECHNOLOGY | 3MT03 LESSON #4: SPECIMEN COLLECTION, CULTURE AND INOCULATION TECHNIQUE Stool (Gl Infection) Throat Swab Collection - tonsillar arches and the o Examples of types of microorganisms you posterior nasopharynx, without touching the sides of the want to asses in stool culture sample: mouth) ▪ Helicobacter pylori Nasal Swab Collection - right and left nostrils and rub ▪ Campylobacter jejuni firmly against the turbinate; use 2 swabs) ▪ Vibrio Cholerae ▪ Member of Enterobacteriaceae SPECIMEN LABELING o Rectal swab (3 specimen) "Do not ACCEPT unlabeled specimen" o Transport medium (Cary blair, Stuart or Specimen Label Amies) o Name Sputum (Pneumonia) o Identification number o Causative agent of Pneumonia and o Room number Tuberculosis: o Physician ▪ Klebsiella pneumoniae o Culture site ▪ Streptococcus pneumoniae o Date of collection ▪ Mycobacterium tuberculosis o Time of collection o Rinse the mouth with water Requisition Form ▪ Do not use mouthwash. o Patient's name o Expectorated sputum o Patient's age (or date of birth) and gender ▪ Deep coughing sputum. o Patient's room number or location o Induced sputum o Physician's name, address, and phone ▪ Uses a device that is injected with number an aerosol material and it will o Specific anatomic site induced your sputum. o Date and time of specimen collection o To rule out Mycobacterium tuberculosis and o Clinical diagnosis or relevant patient history Fungal Infection = 3 separate early o Antimicrobial agents (if patient is receiving morning specimen must be submitted on any) the same day. o Name of individual transcribing order Delta checking - Reviewing a patient's medical history. o Direct Sputum Smear Microscopy Table No. 2 Specimen Storage Guidelines ▪ To scan and screen if the SPECIMEN STORAGE GUIDELINES sputum is suitable specimen for REFRIGERATE ROOM TEMPERATURE culture. Catheter tips (IV) Abscess, lesion, wound CSF for vruses Body fluids CSF CSF for bacteria Feces (unpreserved) Ear: inner Feces for Clostiridium difficile Feces (preserved) toxin (up to 3 days; >3 days store at -70°C) Sputum Genital Urine (unpreserved) Nasal, N/P, throat Tissue Urine (preserved) CSF, Cerebrospinal fluid; IV, intravenous; NIP, nasopharynx TAKE NOTE: SPECIMEN TRANSPORTATION Ideally specimen is transported within 30 mins (No All specimen must be leak proof fixative) Triple Packaging technique For anaerobic bacteria, transport should not take more o Primary, Secondary and Tertiary receptacle than 10 minutes International Air Transport Association (IATA) and CSF Samples (2nd tube, because it is sterile and ideal for International Civil Aviation Organization (ICAO) microbiologic analysis) within 15 minutes o Requirement must be met prior to shipment of biological specimen 3 TRINITY UNIVERSITY OF ASIA – COLLEGE OF MEDICAL TECHNOLOGY | 3MT03 LESSON #4: SPECIMEN COLLECTION, CULTURE AND INOCULATION TECHNIQUE SPECIMEN REJECTION The information on the requisition does not match the information on the specimen label. If the patient name or source does not match, the specimen should be collected again. There is no patient identification on specimen container. The specimen is not submitted in the appropriate transport container or the container is leaking. The quantity of the specimen is inadequate to perform all tests requested. The specimen transport time is more than 2 hours and the specimen has not been preserved. The specimen is received in a fixative such as formalin; stools for O & P examinations are an exception An anaerobic culture is requested on a specimen in which anaerobes are indigenous. Microbiology processing of a particular specimen results in questionable data (e.g., Foley catheter tip). The specimen is dried up. PRESERVATIVES AND ANTICOAGULANT More than one specimen from the same source was Preservative submitted from the same patient on the same day; blood o Boric acid cultures are an exception. o Formalin and Polyvinyl alcohol (Troph and One swab was submitted with multiple requests for Cyst) various organisms. o Refrigeration/Freezing (C. difficille toxin) Gram stain of expectorated sputum reveals fewer than Anticoagulant 25 white blood cells (WBCs) and more than 10 o Sodium Polyanethol Sulfonate (0.025%) epithelial cells per low-power field and mixed bacterial o Heparin (Mycobacterium tuberculosis) flora ▪ TB Quantiferon - Blood examination for Mycobacterium SPUTUM CRITERIA tuberculosis Bartlett's Q scoring of Sputum Samples ▪ Quantiferon tube (3 tubes) - Alternative is Lithium Heparinized Table No. 4 Bartlett's Q scoring of Sputum Samples tube BARLETT’S Q SCORING OF SPUTUM SAMPLE o Citrate and EDTA not used in Microbiology Score Average number Score Average number of ▪ Because it is toxic in microorganism of neutrophils/LPF Squamous ▪ Mechanism action: Chelates Epithelial Cells /LPF Calcium. 0 0 (none) 0 0 (none) +1 1-9 (few) +1 1-9 (few) SPECIMEN PRIORITIZATION +2 10-24 (moderate +2 10-24 (moderate numbers) numbers) Table No. 3 Levels of Specimen Prioritization +3 ≥ 25 (many, +3 ≥ 25 (many, LEVELS OF SPECIMEN PRIORITIZATION numerous) numerous) LEVEL Description Specimens Interpretation: 1 Critical/ Amniotic Fluid Q score: Points average number of neutrophils + invasive Blood POints for average number of squamous cells Cerebrospinal fluid ○ Minimum score: -3 Heart valves ○ Maximum score: +3 Pericardial fluid The higher the score, the better the specimen. 2 Unpreserved Body fluids (noy listed A specimen witha composite score ≥ +1 should be for level 1) cultured. Bone A sputum specimen from a leukopenic patient with Drainage from wounds ciliated respiratory epithelial cells should be cultured. Feces Sputum Tissue 3 Quantitation Catheter tip required Urine Tissue for quantitation 4 Preserved Feces in preservative Urine in preservative Swabs in holding medium (aerobic and anaerobic) 4 TRINITY UNIVERSITY OF ASIA – COLLEGE OF MEDICAL TECHNOLOGY | 3MT03  LESSON #4: SPECIMEN COLLECTION, CULTURE AND INOCULATION TECHNIQUE Murray-Washington Method for Contamination Assessment Table No. 5 Murray-Washington Method for Contamination Assessment MURRAY-WASHINGTON METHOD FOR CONTAMINATION ASSESSMENT Group Epithelial Cells/ Group Leukocytes /LPF LPF 1 >25 1 25 2 10-25 3 >25 3 >25 4 10-25 4 >25 5 25 Interpretation: Ideally, only samples that fall unders groups 4 and 5 are suitable for culture. However, immune suppressed individuals will have reduced numbers of leukocytes in their secretions as well when their cell counts diminish. Therefore, teh criteria have been revised so that the number of epithelial cells (>25 per low-power field) is a b etter indicator of mucosal or saliva contamination. REFERENCES 1. Notes from the discussion by SIR NATHANIEL VERON M. CASAMINA, MSMT, RMT, ASCP 5 TRINITY UNIVERSITY OF ASIA – COLLEGE OF MEDICAL TECHNOLOGY | 3MT03