Simultaneous detection of eleven sexually transmitted agents using multiplexed PCR coupled with MALDI-TOF analysis.
MALDI-TOF mass spectrometry
digital PCR
high-throughput
sexually transmitted infections
timely diagnosis
Journal
Infection and drug resistance
ISSN: 1178-6973
Titre abrégé: Infect Drug Resist
Pays: New Zealand
ID NLM: 101550216
Informations de publication
Date de publication:
2019
2019
Historique:
received:
15
06
2019
accepted:
10
08
2019
entrez:
8
11
2019
pubmed:
7
11
2019
medline:
7
11
2019
Statut:
epublish
Résumé
Sexually transmitted infections (STIs), representing a major global health problem, are caused by different microbes, including bacteria, viruses, and protozoa. Unfortunately, infections of different sexually transmitted pathogens often present similar clinical symptoms, so it is almost impossible to distinguish them clinically. Therefore, the aim of the current study was to develop a sensitive, multitarget, and high-throughput method that can detect various agents responsible for STIs. We developed and tested a 23-plex PCR coupled with matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) assay (sexually transmitted infection-mass spectrometry, STI-MS) that simultaneously targets 11 different agents, including 8 most common clinical pathogens related to STIs (HSV-1, HSV-2, The results showed that the STI-MS approach can accurately detect the expected agents, without cross-reaction with other organisms. The limit of detection of each STI-MS assay was ranged from 1.739 to 10.009 copies/reaction, using probit analyses. The verification rate for each target organism of the STI-MS ranged from a minimum of 89.3% to a maximum of 100%, using conventional assays and ultrasensitive digital PCR to confirm the STI-MS-positive results. To further evaluate the clinical performance of this assay, 241 clinical specimens (124 urethral/cervical swabs and 117 urine) were tested in parallel using the STI-MS assay and monoplex real-time PCR for each agent. The overall validation parameters of STI-MS were extremely high including sensitivity (from 85.7% to 100%), specificity (from 92.3% to 100%), PPV (from 50% to 100%), and NPV (from 99.1% to 100%) for each target. STI-MS is a useful high-throughput screening tool for detecting mixed infections of STIs and has great potential for application in large-scale epidemiological programs for specific microorganisms of STI.
Identifiants
pubmed: 31695443
doi: 10.2147/IDR.S219580
pii: 219580
pmc: PMC6717854
doi:
Types de publication
Journal Article
Langues
eng
Pagination
2671-2682Informations de copyright
© 2019 Xiu et al.
Déclaration de conflit d'intérêts
Junping Peng, Junhua Guo, and Leshan Xiu have a patent (201910527364.8, People’s Republic of China) related to this article. The authors have declared that they have no other conflicts of interest in this work.
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