title: Sensitive simultaneous detection of seven sexually transmitted agents in semen by multiplex-PCR and of HPV by single PCR.
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.
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: Fabrícia
authors.initials: F
authors.lastname: Gimenes
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: Fabiana Soares
authors.initials: FS
authors.lastname: Medina
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: André Luelsdorf Pimenta de
authors.initials: AL
authors.lastname: Abreu
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: Mary Mayumi Taguti
authors.initials: MM
authors.lastname: Irie
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: Isis Baroni
authors.initials: IB
authors.lastname: Esquiçati
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: Natália
authors.initials: N
authors.lastname: Malagutti
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: Vinícius Rodrigo Bulla
authors.initials: VR
authors.lastname: Vasconcellos
authors.affiliation: Clinical Chemistry and Toxicology Department, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Michele Garcia
authors.initials: MG
authors.lastname: Discacciati
authors.affiliation: College of Medicine, Department of Pharmacology, University of Illinois at Chicago, Illinois, Chicago, United States of America.
authors.forename: Marcelo Gialluisi
authors.initials: MG
authors.lastname: Bonini
authors.affiliation: Clinical Chemistry and Toxicology Department, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Silvya Stuchi
authors.initials: SS
authors.lastname: Maria-Engler
authors.affiliation: Section of Clinical Cytology, Department of Clinical Analysis and Biomedicine, State University of Maringá, Maringá, Paraná, Brazil.
authors.forename: Marcia Edilaine Lopes
authors.initials: ME
authors.lastname: Consolaro
mesh_terms: D000328:Adult; D052685:Alphapapillomavirus / Q000302:isolation & purification; D002692:Chlamydia trachomatis / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D045704:Mycoplasma genitalium / Q000302:isolation & purification; D009344:Neisseria gonorrhoeae / Q000302:isolation & purification; D012661:Semen / Q000382:microbiology* / Q000821:virology; D012680:Sensitivity and Specificity; D012749:Sexually Transmitted Diseases / Q000175:diagnosis* / Q000382:microbiology / Q000821:virology; D018139:Simplexvirus / Q000302:isolation & purification; D014208:Treponema / Q000302:isolation & purification