A multiplex PCR assay for the simultaneous detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis

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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Design of a multiplex PCR assay for the simultaneous detection and confirmation of Neisseria gonorrhoeae

Journal of Clinical Pathology ◽

10.1136/jcp.2009.071431 ◽

2010 ◽

Vol 63 (5) ◽

pp. 431-433 ◽

Cited By ~ 1

Author(s):

Isabelle O'Callaghan ◽

Daniel Corcoran ◽

Brigid Lucey

Keyword(s):

Neisseria Gonorrhoeae ◽

Multiplex Pcr ◽

Internal Control ◽

Limit Of Detection ◽

Simultaneous Detection ◽

Pcr Primers ◽

Real Potential ◽

Pcr Assay ◽

Neisseria Species ◽

Multiplex Pcr Assay

ObjectivesTo improve the detection of Neisseria gonorrhoeae by designing a multiplex PCR assay using two N gonorrhoeae-specific genes as targets, thereby providing detection and confirmation of a positive result simultaneously.MethodsPCR primers were designed to detect two N gonorrhoeae genes, namely porA and pgi1. Primers for an internal control targeting the ompW gene of Vibrio cholerae were also designed and incorporated in the assay. The DNA of 45 clinical isolates including 33 N gonorrhoeae isolates, seven non-gonococcal Neisseria species, and five non-Neisseria species was tested using the multiplex PCR assay.ResultsAll 33 N gonorrhoeae isolates were successfully detected by the assay and none of the non-gonococcal isolates was detected. The assay showed a sensitivity and specificity of 100%, and a limit of detection of 1.25 ng of DNA.ConclusionThis multiplex PCR assay offers a sensitive and specific assay suitable for the detection of N gonorrhoeae, and offers real potential for diagnostic use.

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Development and evaluation of a multiplex PCR assay for simultaneous detection ofFlavobacterium psychrophilum,Yersinia ruckeriandAeromonas salmonicidasubsp.salmonicidain culture fisheries

Journal of Veterinary Science ◽

10.4142/jvs.2010.11.3.235 ◽

2010 ◽

Vol 11 (3) ◽

pp. 235 ◽

Cited By ~ 6

Author(s):

Ertan Emek Onuk ◽

Alper Ciftci ◽

Arzu Findik ◽

Yuksel Durmaz

Keyword(s):

Multiplex Pcr ◽

Simultaneous Detection ◽

Pcr Assay ◽

Multiplex Pcr Assay

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A new one-step multiplex PCR assay for simultaneous detection and identification of avian haemosporidian parasites

Parasitology Research ◽

10.1007/s00436-018-6153-7 ◽

2018 ◽

Vol 118 (1) ◽

pp. 191-201 ◽

Cited By ~ 17

Author(s):

Arif Ciloglu ◽

Vincenzo A. Ellis ◽

Rasa Bernotienė ◽

Gediminas Valkiūnas ◽

Staffan Bensch

Keyword(s):

Multiplex Pcr ◽

Simultaneous Detection ◽

Pcr Assay ◽

Haemosporidian Parasites ◽

Multiplex Pcr Assay ◽

Detection And Identification ◽

One Step

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Multiplex PCR Detection of Enterotoxin Genes in Aeromonas spp. from Suspect Food Samples in Northern Taiwan

Journal of Food Protection ◽

10.4315/0362-028x-71.10.2094 ◽

2008 ◽

Vol 71 (10) ◽

pp. 2094-2099 ◽

Cited By ~ 14

Author(s):

YU-CHANG CHANG ◽

JAN-YI WANG ◽

AMMAIYAPPAN SELVAM ◽

SHU-CHEN KAO ◽

SHANG-SHYNG YANG ◽

...

Keyword(s):

Multiplex Pcr ◽

Diarrheal Disease ◽

Simultaneous Detection ◽

Food Poisoning ◽

Food Samples ◽

Pcr Assay ◽

Multiplex Pcr Assay ◽

Enterotoxin Genes ◽

Causative Agents ◽

Northern Taiwan

Aeromonads possess an array of virulence factors and are causative agents of a number of human infections. Among them, genes of one cytotoxic (Act) and two cytotonic (Alt, Ast) enterotoxins are implicated in a human diarrheal disease. A rapid, specific, simultaneous detection of these enterotoxin genes in suspected food poisoning samples is not yet reported. Hence, a multiplex PCR assay was designed to amplify the cytotoxic (act), heat-labile cytotonic (alt), and heat-stable cytotonic (ast) enterotoxin genes of aeromonads. The PCR assay was tested with 133 Aeromonas spp. isolated from suspect food poisoning samples and retail samples of poultry and fish from wet markets in and around Taipei, Northern Taiwan. The Aeromonas spp. isolates were divided into six genotypes based on absence or presence of one or more enterotoxin genes. Of these 133 isolates, Aeromonas caviae (52.5%) and Aeromonas hydrophila (43.4%) were the most frequently isolated species from food poisoning samples and retail samples, respectively. Among the species, A. hydrophila had a significantly higher proportion for harboring three enterotoxin genes than had the others, whereas Aeromonas encheleia, considered a nonpathogen, was found harboring three enterotoxin genes. The multiplex PCR assays are rapid and specific, and provide a useful tool for the detection and genotyping of enterotoxin genes of aeromonads.

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