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Use of a dual priming oligonucleotide system to detect multiple sexually transmitted pathogens in clinical specimens
Author(s) -
Horii T.,
Ohtsuka H.,
Osaki M.,
Ohkuni H.
Publication year - 2009
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2009.02618.x
Subject(s) - virology , biology , priming (agriculture) , oligonucleotide , sexually transmitted disease , microbiology and biotechnology , genetics , human immunodeficiency virus (hiv) , gene , syphilis , botany , germination
Aims: To evaluate a new dual priming oligonucleotide (DPO)‐based multiplex polymerase chain reaction (PCR) assay for detection of six sexually transmitted pathogens, including Chlamydia trachomatis , Neisseria gonorrhoeae , Mycoplasma genitalium , Mycoplasma hominis , Ureaplasma urealyticum and Trichomonas vaginalis . Methods and Results: Using 130 clinical specimens, the results obtained by the multiplex PCR, previously established in‐house PCR and COBAS Amplicor PCR assays were compared. The specimens frequently contained multiple pathogens (34/130 specimens). The multiplex PCR assay had an overall sensitivity of 96% and specificity of 100% compared to the in‐house PCR assay at >20 μg ml −1 of DNA concentrations in samples and there was no cross‐reaction with nonpathogenic Neisseria species that cause the majority of false‐positive results with the COBAS Amplicor PCR assay. Conclusions: The DPO‐based multiplex PCR assay detected the six sexually transmitted pathogens in clinical specimens with a high sensitivity and specificity, although its sensitivity was dependent on the DNA content of the samples. Significance and Impact of the Study: It is the first report about the new DPO‐based technique to detect multiple sexually transmitted pathogens in a single assay, which has considerable potential to diagnose the infections accurately and rapidly.