Multiplex PCR for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Genitourinary specimens

Mahony, James B.; Luinstra, Kathy; Tyndall, Mark; Sellors, John; Krepel, J; Chernesky, Max

doi:10.1128/jcm.33.11.3049-3053.1995

Cited by 68 publications

(27 citation statements)

References 28 publications

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“…Multiplex PCRs capable of detecting 13 or more separate regions of chromosomal DNA and of detecting and typing several bacterial pathogens have been described previously (5,20). However, the use of multiplex RT-PCR for the detection of multiple pathogens with RNA genomes has been much more limited, possibly due to the difficulties of overcoming the inherent inefficiency of the RT step in the RT-PCR or of nucleic acid extraction when the starting material is of poor quality.…”

Section: Discussionmentioning

confidence: 99%

Multiplex PCR for Typing and Subtyping Influenza and Respiratory Syncytial Viruses

et al. 1998

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A multiplex reverse transcription (RT)-PCR method that has been developed is capable of detecting and subtyping influenza A (H1N1 and H3N2) and B viruses as well as respiratory syncytial virus (RSV) types A and B in respiratory clinical samples taken as part of a national community-based surveillance program of influenza-like illness in England and Wales. The detection of each different pathogen depended on distinguishing five amplification products of different sizes on agarose gels following RT-PCR with multiple primer sets. The multiplex RT-PCR was tested with 65 nasopharyngeal apirates from which RSV had been isolated and 237 combined nose and throat swabs from which influenza A (H1N1 and H3N2) or B virus had been detected by virus isolation, as well as 40 respiratory samples from which other viruses including cytomegalovirus, herpes simplex virus, enteroviruses, and parainfluenza viruses had been grown. For the typing and subtyping of influenza A and B viruses and RSV types A and B, the multiplex RT-PCR gave an excellent (100%) correlation with the results of conventional typing and subtyping with specific antisera. Multiplex RT-PCR can also be used to accurately detect more than one viral template in the same reaction mixture, allowing viral coinfections to be identified with the same respiratory specimen.

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Section: Discussionmentioning

confidence: 99%

Multiplex PCR for Typing and Subtyping Influenza and Respiratory Syncytial Viruses

et al. 1998

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“…A multiplex PCR (primary m-PCR) with primers KL-1/ KL-2 (targeting 241 bp fragment of the cryptic plasmid in C. trachomatis) and HO1/ HO3 (targeting 390 bp fragment of the cppB gene in N. gonorrhoeae) was performed to detect both organisms in one reaction tube, as described previously (13). Prior to testing of clinical specimens, PCR conditions were optimized as uniplex PCR using the DNA extracts from standard reference strains of C. trachomatis (serovar L2 type strain 434/Bu, ATCC VR-902B) and N. gonorrhoeae (ATCC 11688).…”

Section: Pcr Assaymentioning

confidence: 99%

Transmission Rates of Chlamydia trachomatis and Neisseria gonorrhoeae Infections from Pregnant Women to Newborns, Tehran, Iran

Esteghamati

Mazouri

Sayyahfar

et al. 2020

Jundishapur J Microbiol

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Background: Pregnant women with Chlamydia trachomatis and Neisseria gonorrhoeae infections can vertically transmit these microorganisms to their newborns through the birth canal and cause neonatal conjunctivitis secondary to sexually transmitted infections. Objectives: In this cross-sectional study, we aimed to evaluate the prevalence of C. trachomatis and N. gonorrhoeae infections among pregnant women attending a hospital in Tehran, and also determine the vertical transmission rate of these two organisms to the eyes of newborns after vaginal delivery. Methods: Endocervical and conjunctival swabs were collected from pregnant women and their newborns within 24 hours after birth. Demographic and clinical data of participants were obtained using a questionnaire and from the hospital records. Then, DNA was extracted and tested by a multiplex PCR assay to detect C. trachomatis and N. gonorrhoeae in specimens. Results: Genital infections of C. trachomatis and N. gonorrhoeae were detected in 9.6% (11 of 125) and 1.6% (2 of 125) of pregnant women, respectively. Among newborns, ocular infection with C. trachomatis was detected in 2 (1.6%), and no case of N. gonorrhoeae infection was found. Both infected infants were born from asymptomatic infected women. Therefore, the vertical transmission rate of C. trachomatis infection was calculated as 18.1%. Our results also revealed that ocular C. trachomatis infection in neonates is significantly in association with genital C. trachomatis infection in pregnant women (OR = 0.16, 95% CI = 0.03 -0.7, P = 0.002). Conclusions: Pregnant women with asymptomatic infection of C. trachomatis have a key role in the distribution of chlamydial conjunctivitis in newborns. Since ocular prophylaxis in neonates is not effective for chlamydial conjunctivitis, therefore education and screening of pregnant women, as well as treatment of infected cases, remain as the best approach for controlling the disease.

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“…Mahony and colleagues developed a multiplex PCR assay for the simultaneous detection of C. trachomatis and N. gonorrhoeae in first void urine specimens. They reported the sensitivity of multiplex PCR for detecting C. trachomatis was 100%, compared with 81.8% for enzyme immunoassay and for detecting N. gonorrhoeae was 92.3% compared with 84.6% for urethral culture and the specificity was 100% for both C. trachomatis and N. gonorrhoeae [24]. In comparison, our multiplex PCR has a similar sensitivity/specificity for detection of C. trachomatis but higher sensitivity for detection of N. gonorrhoeae on endocervical specimens.…”

Section: Discussionmentioning

confidence: 65%

A multiplex assay of Trichomonas vaginalis, Chlamydia trachomatis and Neisseria gonorrhoeae infections in genital specimens

Rostami

Rashidi

Nazari

et al. 2017

J Infect Dev Ctries

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Introduction: A significant proportion of patients with Sexually Transmitted Infections (STIs) are coinfected with multiple pathogens. We report here development of a multiplex PCR for simultaneous detection of Chlamydia trachomatis (C.trachomatis), Neisseria gonorrhoeae (N.gonorrhoeae) and Trichomonas vaginalis (T.vaginalis) in genital specimens from women. Methodology: After detection of the organisms by routine techniques including PCR, culture and direct smear, multiplex-PCR was optimized to detect ompI gene of CT, parC of NG, ITS ribosomal RNA of TV as target genes. The limit of detection (LOD) was determined using serially diluted genomic DNA from known number of each pathogen. Results: Totally 300 volunteers with mean age of 36.5 ±7.03 years were included and 266 (88.7%) had genitourinary clinical manifestations. Of 300 women, 150 (50.0%) were infected. Of them, 17 (5.7%) had N. gonorrhoeae, 98 (32.7%) T. vaginalis and 35 (11.7%) C. trachomatis. Multiplex-PCR revealed a total of 10 coinfections (3.3%) including 2 specimens of C. trachomatis/ N. gonorrhoeae, 3 specimens of C .trachomatis/ T. vaginalis and 5 specimens of N. gonorrhoeae/T. vaginalis coinfections. The sensitivity and specificity of multiplex-PCR for detecting N. gonorrhoeae were 100% and 98.59% (279 of 283) respectively and, for C. trachomatis and T. vaginalis were 100%. The LOD was 0.1 pg of DNA for C. trachomatis and N. gonorrhoeae, and 1.5 pg for T. vaginalis. Conclusions: The performance of this multiplex-PCR makes it a sensitive, rapid and affordable technique in clinical laboratory for simultaneous detection of STIs.

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Multiplex PCR for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Genitourinary specimens

Cited by 68 publications

References 28 publications

Multiplex PCR for Typing and Subtyping Influenza and Respiratory Syncytial Viruses

Multiplex PCR for Typing and Subtyping Influenza and Respiratory Syncytial Viruses

Transmission Rates of Chlamydia trachomatis and Neisseria gonorrhoeae Infections from Pregnant Women to Newborns, Tehran, Iran

A multiplex assay of Trichomonas vaginalis, Chlamydia trachomatis and Neisseria gonorrhoeae infections in genital specimens

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