Use of the mtrR Gene for Rapid Molecular Diagnosis of Neisseria gonorrhoeae and Identification of the Reduction of Susceptibility to Antibiotics in Endocervical Swabs
Neisseria gonorrhoeae is one of the main etiological agents of sexually transmitted diseases. The asymptomatic course of the infection and its resistance to antibiotics can lead to pelvic inflammatory disease and infertility.
We developed a polymerase chain reaction (PCR) test using the methyltetrahydrofolate homocysteine methyltransferase reductase (mtrR) gene to identify N. gonorrhoeae and detect reduced susceptibility to antibiotics.
Material and Methods
We analysed 250 samples of endocervical exudate from infertile women with a negative diagnosis of N. gonorrhoeae. We designed NGmtr primers to detect N. gonorrhoeae and identify the antibiotic-resistant strain.
Of the 250 samples, 60 (24%) tested positive for N. gonorrhoeae using real-time PCR. Our study was validated using the HO primers and the Seeplex STD6 ACE System, with a 100% correlation. Furthermore, the NGmtr primers are specific for N. gonorrhoeae and not for other species. Additionally, the curves generated by real-time PCR differed between wild and variant strains (10.93%). The dissociation temperatures for the wild and variant strains were 86.5 and 89 °C, respectively.
The NGmtr primers enabled us to identify N. gonorrhoeae strains with or without reduction of susceptibility to antibiotics. Therefore, this work constitutes a tool that will facilitate the diagnosis of this infection for a low cost and improve patient quality of life.
The authors thank the Institute of Biology of the UNAM for the sequencing service provided for this work.
Compliance with Ethical Standards
Conflict of interest
Marcos R. Escobedo-Guerra, Mitzuko Katoku-Herrera, Marcela Lopez Hurtado, Rodrigo Gutierrez Trujillo, and Fernando M. Guerra-Infante are researchers from the National Institute of Perinatology and have no conflicts of interest that are directly relevant to the content of this work.
This work was funded by the National Institute of Perinatology.
Ethical approval and informed consent
This work was regulated by the Ethics Committee of the National Institute of Perinatology (Approval number: 212250-3120-10607-01-14).
- 1.Newman L, Rowley J, Vander Hoorn S, Wijesooriya NS, Unemo M, Low N, et al. Global estimates of the prevalence and incidence of four curable sexually transmitted infections in 2012 based on systematic review and global reporting. PLoS One. 2015;10(12):e0143304.CrossRefPubMedPubMedCentralGoogle Scholar
- 3.Detels R, Green AM, Klausner JD, Katzenstein D, Gaydos C, Handsfield HH, et al. The incidence and correlates of symptomatic and asymptomatic Chlamydia trachomatis and Neisseria gonorrhoeae infections in selected populations in five countries. Sex Transm Dis. 2011;38(6):503–9.PubMedPubMedCentralGoogle Scholar
- 8.Whiley DM, Limnios A, Moon NJ, Gehrig N, Goire N, Hogan T, et al. False-negative results using Neisseria gonorrhoeae porA pseudogene PCR—a clinical gonococcal isolate with an N. meningitidis porA sequence, Australia, March 2011. Euro Surveill Bull Eur Sur Mal Transm Eur Commun Dis Bull. 2011;16(21).Google Scholar
- 18.Colson P, Gouriet F, Badiaga S, Tamalet C, Stein A, Raoult D. Real-time laboratory surveillance of sexually-transmissible infections in Marseille University hospitals reveals rise of gonorrhoea, syphilis and human immunodeficiency virus seroconversions in 2012. Euro Surveill Bull Eur Sur Mal Transm Eur Commun Dis Bull. 2013;18(7):4.Google Scholar
- 24.van Bergen JE, Spaargaren J, Götz HM, Veldhuijzen IK, Bindels PJ, Coenen TJ, et al. Population prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae in the Netherlands. Should asymptomatic persons be tested during Population-based Chlamydia Screening also for gonorrhoea or only if chlamydial infection is found? BMC Infect Dis. 2006;6:42.CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Vernel-Pauillac F, Nandi S, Nicholas RA, Goarant C. Genotyping as a tool for antibiotic resistance surveillance of Neisseria gonorrhoeae in New Caledonia: evidence of a novel genotype associated with reduced penicillin susceptibility. Antimicrob Agents Chemother. 2008;52(9):3293–300.CrossRefPubMedPubMedCentralGoogle Scholar