A New Multiplex Real-time PCR Test for HSV 1 / 2 and Syphilis- An evaluation of its impact in the laboratory and clinical setting.
Résumé
Objectives: To develop, evaluate and implement a new multiplex real-time PCR test for the detection of HSV-1, HSV-2 and syphilis. Methods: A new real-time PCR assay for syphilis was developed and multiplexed with an existing assay for the detection of HSV-1 and 2. This new multiplex assay was validated using sensitivity and specificity panels and external quality assurance (EQA) material. In the first 6 months after clinical launch, 692 samples were taken from clients attending with suspected HSV or primary syphilis. DNA was extracted using the virus kit on the MDX extractor (Qiagen) and PCR performed on an ABI 7500 system (Applied Biosystems). Syphilis PCR positives were confirmed by a second PCR at the national reference laboratory. PCR results were aligned with dark ground microscopy (DGM) findings and serological tests for syphilis where taken. Results: The sensitivity and specificity panels showed the test to be reliable and the EQA panels showed the test to be robust. Of the 692 samples, 139 were positive for HSV-1, 136 for HSV-2, 15 for syphilis, one for both syphilis and HSV-1, and 401 were negative. The reference laboratory confirmed all syphilis PCR positive samples. The PCR test was more sensitive than both DGM and serological testing for the diagnosis of primary syphilis and had positive and negative predictive values of 100%. Conclusions: The introduction of this new test has led to a better turn around time for the diagnosis of genital ulcer disease, better detection of primary syphilis infection and the detection of unexpected cases of syphilis where the aetiological agent suspected was HSV.
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