Molecular Method Identifies Six Sexually Transmitted Pathogens

A multiplex polymerase chain reaction (PCR)-based auto-capillary electrophoresis (ACE) system can distinguish six pathogens that are transmitted during sexual contact.

The method combines a nucleic acid amplification technique with a fully automated electrophoretic device and was developed to simplify the analysis of multiplex PCR assays and to provide accurate amplicon sizing with automated interpretation of pathogen identity.

Scientists at the Rabin Medical Center, (Petach Tikva, Israel), screened genital/urinary specimens collected from 113 patients, 72 male and 41 female, with a sexually transmitted infection. The pathogens investigated were: Chlamydia trachomatis, Mycoplasma hominis, M. genitalium, Neisseria gonorrhoeae, Trichomonas vaginalis, and the genital Ureaplasma, (U. urealyticum and U. parvum). The method used a dual priming oligonucleotide (DPO) PCR system, the Seeplex STD6B ACE. The amplified PCR products were separated and detected by the TapeStation automated gel electrophoresis-detection system. The results were compared with conventional reference methods, including culture and PCR tests.

The sensitivity of STD6B ACE assay was found to be 100% for five of the pathogens and 98% for genital Ureaplasma. A single pathogen was detected in 51 specimens, and two or more pathogens were detected in 24 samples. The specificity of the assays ranged from 97% to 100%. The most prevalent pathogen overall was Ureaplasma, which was identified in 42.5% specimens. N. gonorrhoeae was eight-fold more prevalent in men than in women, whereas M. hominis was more prevalent in women, 21.9% versus 5.6% in men. The automated Seeplex STD6B ACE multiplex PCR is a product of Seegene (Seoul, Korea), and the automated gel electrophoresis system TapeStation, is manufactured by Lab901 Ltd. (Bilston Glen, Loanhead, UK).

The authors concluded that the multiplex PCR and ACE system is highly sensitive and specific for the rapid, simultaneous detection of sexually transmitted pathogens directly from a single specimen. The STD6B ACE system uses prepackaged reagents and is rationally automated, which simplifies the analysis of multiplex PCR assays, eliminating the need for time- and labor-consuming agarose gel electrophoresis and thereby improving laboratory workflow. Application of the system may facilitate the diagnosis, treatment, and care of patients with sexually transmitted diseases and help to prevent the spread of these infections. The study was published in May 2011 in the journal Diagnostic Microbiology and Infectious Disease.

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Rabin Medical Center
Seegene
Lab901 Ltd.