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MDCH STATE LABORATORY TUBERCULOSIS TESTING Specimens/Tests/Reports Dale E. Berry, B.S. TB Lab Manager (517) 335-9637

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Presentation on theme: "MDCH STATE LABORATORY TUBERCULOSIS TESTING Specimens/Tests/Reports Dale E. Berry, B.S. TB Lab Manager (517) 335-9637"— Presentation transcript:

1 MDCH STATE LABORATORY TUBERCULOSIS TESTING Specimens/Tests/Reports Dale E. Berry, B.S. TB Lab Manager (517)

2 MDCH STATE LABORATORY LANSING, MICHIGAN

3 TYPES OF SPECIMENS SPUTUM PULMONARY ASPIRATES GASTRIC ASPIRATES BODY FLUIDS TISSUES BLOOD FECES OTHER

4 SPECIMEN QUALITY Three specimens on three successive days Early morning specimens (First thing out!) Adequate specimen volume (>7 ml) Sputum not saliva (epithelial cells?) Condition of specimen (pH, etc.) Appropriate specimen container Complete and accurate information Proper packaging (i.e. US Postal Service) “Prompt” delivery to MDCH

5 MDCH – TESTS for AFB AFB Slide Examination Direct Amplified Genetic Probe (M.tb complex) Direct Susceptibility AFB Culture and Isolation Pre-Identification (Genetic Probe/HPLC) Final Identification (Biochemical, etc.) Pre-Indirect Susceptibility (Primary Drugs(1 st Line)) Final-Indirect M.tb. Susceptibility (2 nd /3 rd Line) MOTT Susceptibility DNA Fingerprinting

6 AFB Slide Examination Least Sensitive AFB Test Requires 100,000 AFB/ml Requires an adequate specimen Negative smear – Does not rule out AFB Positive smear – AFB present (viability?) AFB Positive = Mycobacterium spp., etc. Help support TB diagnosis Quantitation: Positive/Positive Few/ Negative

7 MTD – Direct Amplified Probe Rapid Test & Sensitive Specific – M.tuberculosis complex Not a test of viability or cure Pulmonary Specimens only Specimen MDCH only Reduces subsequent test sensitivity Delays culture & susceptibility results

8 AFB Culture and Isolation More sensitive than slide Requires only 10 AFB/ ml of specimen AFB Growth in 1-2 weeks (Bactec) Genetic Probes and HPLC Rapid ID Fewer AFB/ml – Delayed growth “AFB Growth” required for other tests Mixed cultures require subcultures

9 Preliminary AFB Identification Genetic Probe/HPLC One day after culture positive Probe for M.tuberculosis complex HPLC for MOTT Helps confirm antibiotic therapy

10 Preliminary TB Susceptibility Approximately 1 week after pre-ID Tests “Primary” anti-Tuberculosis drugs Direct and Indirect testing Bactec methodology Confirmed by“Proportion Plate Method”

11 Final AFB Identification Required to differentiate M.tb complex Required to confirm HPLC ID’s Required to clarify unresolved ID’s Requires additional time (3-4 weeks) Final Report ASAP

12 TB Susceptibility - Additional Proportion Plate Method Requires additional 3 weeks To increases choices for therapy Performed for drug resistant TB cases Tests 2 nd and 3 rd line antibiotics Confirms results of “Broth Method”

13 MOTT Susceptibility Testing Test according to latest NCCLS Guidelines M.avium complex - Clarithromycin M.kansasii/M.marinum - Rifampin/Ethambutol M.fortuitum/M.chelonae – Refer to CDC Additional antibiotics not offered by MDCH Refer to National Jewish Hospital in Denver, Colorado

14 RFLP – DNA Fingerprinting For TB outbreak investigations Check for “reactivation vs re-infection Check for lab cross-contamination Performed on approved requests only

15 MDCH AFB TEST TIME-LINE SPECIMENMTDCULTURETB SENSFINAL ID FINAL CULTSLIDE 24 HRS 48 HRS7-10 DA 8-14 DA DA3 WKS6 WKS AFB FOUND NOT FOUND TB complex RNA DETECTED NOT DETECTED MYCOBACTERIUM FOUND PRELIM ID M.tb complex NOT M.tb complex MOTT SUSCEPTIBILITY PRIMARY ANTIBIOTICS FINAL ID SPECIES CONFIRMATION MYCOBACTERIUM NOT FOUND REFERRED CULTURE24-48 HRS7-10 DA2-3 WKS PRE ID PROBE HPLC

16 QUESTIONS > CALL MDCH TB Laboratory Dale Berry Lab (517) Steve Church Microbiologist Angie Schooley Microbiologist Jolene Vanneste (517)

17 TB CONTACT TB PATIENT SYMPTOMS HEALTH CARE PROVIDER (DR) DIAGNOSIS STATE LAB SLIDE MTD CULTURE IDENTIFICATION DNA FINGERPRINT STATE DISEASE CONTROL PRIVATE LAB SLIDE CULTURE PRELIM ID COUNTY HEALTH DEPARTMENT TREATMENT CONTACT INVESTIGATION Complete Networking


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