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Identifying Bacteria Causing Vaccine-Preventable Meningitis

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Identifying Bacteria Causing Vaccine-Preventable Meningitis Identifying Bacteria Causing Vaccine-Preventable Meningitis Sentinel, national, regional, global Health care Clinical specimen laboratories Ministry Vaccination of Health Data management CSF Specimen If specimen cannot be processed in <1 hour, If only one tube, inoculate into Tube microbiology Tube T-I medium. 1 is a priority 3 For details consult Lab Manual or Standard Tube Operating Procedures Chemical analysis: 2 Record overall appearance; protein and glucose tests perform cell count: WBCs Microbiological tests PCR Immunochromatographic Centrifuge Latex Test Agglutination Freeze 250µl-500µl Test of CSF for PCR 100-200µl CSF volume is needed Do not centrifuge if<1ml (if < 500µl CSF When using kits refer When using kits refer to available, freeze to manufacturers manufacturers standard any remaining CSF standard operating procedures (SOPs) operating procedures (SOPs) volume for PCR) in package insert ( Supernatant in package insert Pellet ( Gram Stain Culture on Blood Agar Plate (BAP) and Chocolate Agar Plate (CAP) Decolorize with 95% ethanol; do not decolorize longer than 10 sec. Use sheep or horse blood, NOT human blood S. pneumoniae: H. influenzae: N. meningitidis: BAP CAP BAP or CAP LOOK FOR: gram POS. LOOK FOR: gram NEG. LOOK FOR: gram NEG. diplococci or cocci coccobacilli coffee-bean in short chains shaped diplococci S. pneumoniae H. influenzae N. meningitidis FOR anY otheR GRam stain Results, LOOK FOR: LOOK FOR: LOOK FOR: applY otheR Routine lab methods: Notify clinician of Gram small, grayish, moist (sometimes large, round, colorless- round, moist, glistening, India inK, otheR cultuRE media, etc. stain results within 1 hour! watery) colonies surrounded by to-grey, opaque colonies convex, gray and a greenish zone of α-hemolysis unpigmented colonies Optochin Catalase Growth Factor Kovac’s Oxidase Using optochin (P discs, 5µg, 6mm diameter), Do NOT transfer any of interpret zones for resistance (R) and susceptibility (S) the blood agar to the slide! RBCs in the blood agar will No color change If filter paper turns blue/purple cause a false-positive reaction indicates within a few seconds, indicates negative test presence of either H. influenzae or N. meningitidis Perform additional Gram stain R S on colonies to distinguish organism If no zone (or < 14 mm), If zone of inhibition If bubbling occurs, An absence of Does not require If growth only occurs perform bile solubility is ≥14 mm, indicates not Streptococcus bubbling indicates both hemin (X factor) with BOTH X factor test before ruling out susceptibility and a negative test and NAD (V factor) and V factor: If stain indicates If stain indicates presence of Streptococcus spp. for growth: indicates N. meningitidis, H. influenzae, S. pneumoniae are catalase negative not H. influenzae H. influenzae to confirm,TEST FOR: organism is confirmed Bile Solubility Enter results into surveillance logbook and CTA Sugars If cells do not lyse If cells lyse (dissolve) and report to clinicians and appropriate officials. LOOK FOR: visible turbidity or color change to yellow (insoluble), turbidity disappears, not S. pneumoniae report as S. pneumoniae Link clinical and lab data for each patient Contacts Storage Additional Name Phone Number Tests sucrose, lactose glucose, maltose Hospital surveillance coordinator: ___________________________ ___________________________ Freeze bacterial isolates and any remaining CSF If acid occurs in sucrose If acid production occurs Hospital laboratory coordinator: ___________________________ ___________________________ for molecular testing Antimicrobial susceptibility tests: or lactose, or any other in glucose and maltose, Hospital data manager: ___________________________ ___________________________ at a Reference Lab. to be performed according to pattern of colors appears, but not in sucrose or _______________________________________________________________________________________________ Include label and date on frozen internationally recognized not N. meningitidis lactose, report as Additional information: http://www.who.int/nuvi/surveillance/resources/en/index.html isolates and CSF specimens clinical guidelines N. meningitidis.
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