Annals of Clinical Microbiology, The official Journal of the Korean Society of Clinical Microbiology

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pISSN 2288-0585 eISSN 2288-6850

Comparative Evaluation of Multiplex Real-Time PCR Assays for Six Pathogens of Sexually Transmitted Infections

Original article

Annals of Clinical Microbiology (Ann Clin Microbiol) 2017 March, Volume 20, Issue 1, pages 1-6.

https://doi.org/10.5145/ACM.2017.20.1.1

Comparative Evaluation of Multiplex Real-Time PCR Assays for Six Pathogens of Sexually Transmitted Infections

Hae-Sun Chung, Miae Lee

Department of Laboratory Medicine, Ewha Womans University College of Medicine, Seoul, Korea

Abstract

Background: The multiplex real-time PCR assay is a sensitive test for simultaneous detection of various pathogens of sexually transmitted infections (STIs). We evaluated the performance of two multiplex real-time PCR assays for six STI pathogens.

Methods: DNA samples after being used to conduct PCR for STI pathogens were stored below −70℃. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Trichomonas vaginalis (TV) were detected by multiplex real-time PCR with GeneFinder STD I (CT/NG/UU)/II (MG/MH/TV) Multiplex Real-time PCR Kits (Infopia, Korea; GeneFinder assay) and Real-Q CT&NG/ MH&TV/MG&UU Kits (BioSewoom, Korea; Real-Q assay). Discrepant results were resolved by another multiplex real-time assay, Anyplex II STI-7 Detection (Seegene, Korea). Any two positive results for the assays were considered true positive.

Results: Among 81 samples, the GeneFinder assay detected 63 pathogens from 45 cases (16 CT, 2 NG, 6 MG, 20 MH, 18 UU, and 1 TV) and Real-Q assay detected 66 pathogens from 47 cases (16 CT, 2 NG, 8 MG, 20 MH, 19 UU, and 1 TV). For the results of positive cases and negative cases, the overall concordance rate between the two multiplex real-time assays was 93.8% (Kappa=0.87). For each pathogen, the agreement rates of the two assays ranged from 97.5 to 100% (Kappa>0.8).

Conclusion: There was no significant difference between the results of GeneFinder assay and Real-Q assay. Both multiplex real-time PCR assays can be useful methods for the detection of STI pathogens in clinical laboratories. (Ann Clin Microbiol 2017;20:1- 6)

Keywords

Multiplex real-time polymerase chain reaction, Sexually transmitted infection