Sensitive simultaneous detection of seven sexually transmitted agents in semen by multiplex-PCR and of HPV by single PCR

PLoS One. 2014 Jun 12;9(6):e98862. doi: 10.1371/journal.pone.0098862. eCollection 2014.

Abstract

Sexually transmitted diseases (STDs) may impair sperm parameters and functions thereby promoting male infertility. To date limited molecular studies were conducted to evaluate the frequency and type of such infections in semen Thus, we aimed at conceiving and validating a multiplex PCR (M-PCR) assay for the simultaneous detection of the following STD pathogens in semen: Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, Herpes virus simplex (HSV) -1 and -2, and Treponema pallidum; We also investigated the potential usefulness of this M-PCR assay in screening programs for semen pathogens. In addition, we aimed: to detect human Papillomavirus (HPV) and genotypes by single PCR (sPCR) in the same semen samples; to determine the prevalence of the seven STDs, HPV and co-infections; to assess the possibility that these infections affect semen parameters and thus fertility. The overall validation parameters of M-PCR were extremely high including agreement (99.2%), sensitivity (100.00%), specificity (99.70%), positive (96.40%) and negative predictive values (100.00%) and accuracy (99.80%). The prevalence of STDs was very high (55.3%). Furthermore, associations were observed between STDs and changes in semen parameters, highlighting the importance of STD detection in semen. Thus, this M-PCR assay has great potential for application in semen screening programs for pathogens in infertility and STD clinics and in sperm banks.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Alphapapillomavirus / isolation & purification
  • Chlamydia trachomatis / isolation & purification
  • Humans
  • Male
  • Middle Aged
  • Multiplex Polymerase Chain Reaction / methods*
  • Mycoplasma genitalium / isolation & purification
  • Neisseria gonorrhoeae / isolation & purification
  • Semen / microbiology*
  • Semen / virology
  • Sensitivity and Specificity
  • Sexually Transmitted Diseases / diagnosis*
  • Sexually Transmitted Diseases / microbiology
  • Sexually Transmitted Diseases / virology
  • Simplexvirus / isolation & purification
  • Treponema / isolation & purification

Grant support

This work was supported by grants from Coordenação de Aperfeiçoamento de Pessoal de Nível superior (CAPES), AUX-PE-PRODOC 2571/2010, Brazilian Government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.