Development of Real-Time PCR Methods for the Detection of Bacterial Meningitis Pathogens without DNA Extraction

PloS One
Jeni VuongXin Wang

Abstract

Neisseria meningitidis (Nm), Haemophilus influenzae (Hi), and Streptococcus pneumoniae (Sp) are the lead causes of bacterial meningitis. Detection of these pathogens from clinical specimens using traditional real-time PCR (rt-PCR) requires DNA extraction to remove the PCR inhibitors prior to testing, which is time consuming and labor intensive. In this study, five species-specific (Nm-sodC and -ctrA, Hi-hpd#1 and -hpd#3 and Sp-lytA) and six serogroup-specific rt-PCR tests (A, B, C, W, X, Y) targeting Nm capsular genes were evaluated in the two direct rt-PCR methods using PerfeCTa and 5x Omni that do not require DNA extraction. The sensitivity and specify of the two direct rt-PCR methods were compared to TaqMan traditional rt-PCR, the current standard rt-PCR method for the detection of meningitis pathogens. The LLD for all 11 rt-PCR tests ranged from 6,227 to 272,229 CFU/ml for TaqMan, 1,824-135,982 for 5x Omni, and 168-6,836 CFU/ml for PerfeCTa. The diagnostic sensitivity using TaqMan ranged from 89.2%-99.6%, except for NmB-csb, which was 69.7%. For 5x Omni, the sensitivity varied from 67.1% to 99.8%, with three tests below 90%. The sensitivity of these tests using PerfeCTa varied from 89.4% to 99.8%. The specificity ranges of ...Continue Reading

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Citations

Nov 11, 2017·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Jessica R MacNeilAmanda C Cohn
Nov 4, 2016·Nature Reviews. Disease Primers·Diederik van de BeekEelco Wijdicks
Sep 11, 2019·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Angeline BoulaMartin Antonio
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Sep 11, 2019·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Enyonam TsolenyanuMartin Antonio
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Methods Mentioned

BETA
PCR

Software Mentioned

PerfeCTa

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