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Diagnostics 2018, 8(3), 58; https://doi.org/10.3390/diagnostics8030058

Triplex Real-Time PCR without DNA Extraction for the Monitoring of Meningococcal Disease

1
Centers for Disease Control and Prevention, Atlanta, GA 30329, USA
2
Institut National de Recherche en Santé Publique, Bamako 00223, Mali
3
Centre Hospitalier Universitaire Pédiatrique Charles de Gaulle, Ouagadougou 01, Burkina Faso
4
Instituto Adolfo Lutz, São Paulo 01246-902, Brasil
*
Author to whom correspondence should be addressed.
Received: 30 July 2018 / Revised: 22 August 2018 / Accepted: 26 August 2018 / Published: 30 August 2018
(This article belongs to the Special Issue Diagnosis of Bacterial Pathogens)
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Abstract

Detection of Neisseria meningitidis has become less time- and resource-intensive with a monoplex direct real-time PCR (drt-PCR) to amplify genes from clinical specimens without DNA extraction. To further improve efficiency, we evaluated two triplex drt-PCR assays for the detection of meningococcal serogroups AWX and BCY. The sensitivity and specificity of the triplex assays were assessed using 228 cerebrospinal fluid (CSF) specimens from meningitis patients and compared to the monoplex for six serogroups. The lower limit of detection range for six serogroup-specific drt-PCR assays was 178–5264 CFU/mL by monoplex and 68–2221 CFU/mL by triplex. The triplex and monoplex showed 100% agreement for six serogroups and the triplex assays achieved similar sensitivity and specificity estimates as the monoplex drt-PCR assays. Our triplex method reduces the time and cost of processing CSF specimens by characterizing six serogroups with only two assays, which is particularly important for testing large numbers of specimens for N. meningitidis surveillance. View Full-Text
Keywords: meningococcal meningitis; direct multiplex real-time PCR; Neisseria meningitidis; meningococcal serogroups meningococcal meningitis; direct multiplex real-time PCR; Neisseria meningitidis; meningococcal serogroups
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Whaley, M.J.; Jenkins, L.T.; Hu, F.; Chen, A.; Diarra, S.; Ouédraogo-Traoré, R.; Sacchi, C.T.; Wang, X. Triplex Real-Time PCR without DNA Extraction for the Monitoring of Meningococcal Disease. Diagnostics 2018, 8, 58.

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