Abstract
BackgroundNucleic acid amplification tests (NAAT) are well-accepted in diagnosis and surveillance of sexually infectious pathogens worldwide. However, performance differences between a RNA-based NAAT and DNA-based NAAT are rarely reported. This study compares the performances of the RNA-based SAT (simultaneous amplification and testing) assay and the DNA-based quantitative real-time polymerase chain reaction (qPCR) assay.MethodsA total of 123 urogenital swabs were collected from outpatients with suspected genital infections in our hospital. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Ureaplasma urealyticum (UU) in these swabs were simultaneously tested by SAT and qPCR. Any swabs were positive in the qPCR assay were further verified by following cloning and sequencing. All statistical analysis was performed using the SPSS software.ResultsWhen the concentrations of CT, NG, or UU were more than 1 × 103 copies/ml, 100% agreements between SAT and qPCR were observed regardless of the pathogen. No discrepancy was found. However, the sensitivity of SAT is significantly higher than qPCR in samples with concentration less than 1 × 103 copies/ml. When tested by SAT and qPCR, 57.14 and 28.57% were positive for CT, 46.15% and 0 were positive for NG, 80% and 0 were positive for UU, respectively.ConclusionsThe SAT assay has better agreements and higher sensitivities when compared with the qPCR assay, and thus could be a better choice for screening, diagnosis, and surveillance of sexually transmitted diseases, especially for CT and NG.
Highlights
Nucleic acid amplification tests (NAAT) are well-accepted in diagnosis and surveillance of sexually infectious pathogens worldwide
A novel RNA-based NAAT, which is based on transcription mediated amplification (TMA) and designed to detect pathogenic bacteria by amplification of 16S rRNA, has been reported to detect some sexual, respiratory, and enteric pathogens, including Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), UU, Mycoplasma genitalium, Mycobacterium tuberculosis, Mycoplasma pneumonniae, entervirus 71, and coxsackievirus A16 [7,8,9,10,11]
Of the 123 urogenital swabs, 33 swabs were positive for CT, 32 swabs were positive for NG, and 29 swabs were positive for UU after confirmation by the quantitative real-time polymerase chain reaction (qPCR) assay and sequencing
Summary
Nucleic acid amplification tests (NAAT) are well-accepted in diagnosis and surveillance of sexually infectious pathogens worldwide. Ureaplasma urealyticum (UU) is considered as a commensal bacterium in the urogenital tract, it has been reported to be associated with some sexually transmitted diseases including non-gonococcal urethritis (NGU) [1, 2] and patients with high UU loads are associated with acute NGU [2] These infections are curable, screening still remains very important to prevent late complications caused by these pathogens. Nucleic acid amplification tests (NAAT), which have a better performance with high sensitivity, specificity, and ease of sample transport, have been approved by the Food and Drug Administration (FDA) for the detection of CT and NG, and widely used in clinical facilities to detect the sexually infectious pathogens [5, 6]. Performance comparison of the RNA-based SAT with DNA-based NAAT assay is unclear yet
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