Herpes simplex virus (HSV) and varicella zoster virus (VZV) are related members of the Herpesviridae family and are well-documented human pathogens causing a spectrum of diseases, from mucocutaneous disease to infections of the central nervous system. This study was carried out to evaluate and validate the performance of a multiplex real-time polymerase chain reaction (PCR) assay in detecting and differentiating HSV1, HSV2, and VZV from clinical samples. Consensus PCR primers for HSV were designed from the UL30 component of the DNA polymerase gene of HSV, with 2 separate hydrolysis probes designed to differentiate HSV1 and HSV2. Separate primers and a probe were also designed against the DNA polymerase gene of VZV. A total of 104 clinical samples were available for testing by real-time PCR, conventional PCR, and viral culture. The sensitivity and specificity of the real-time assay was calculated by comparing the multiplex PCR result with that of a combined standard of virus culture and conventional PCR. The sensitivity of the real-time assay was 100%, with specificity ranging from 98% to 100% depending on the target gene. Both PCR methods detected more positive samples for HSV or VZV, compared with conventional virus culture. This multiplex PCR assay provides accurate and rapid diagnostic capabilities for the diagnosis and differentiation of HSV1, HSV2, and VZV infections, with the presence of an internal control to monitor for inhibition of the PCR reaction.

stillbirth, fetal loss, neonatal death, low birth weight, preterm birth and syphilis infection in the infant. In Haiti, 90% of pregnant women report at least one antenatal visit. We evaluated the field performance of the SD BIOLINE HIV/Syphilis Duo test in a high-risk setting in Port-au-Prince, Haiti using whole blood fingerprick specimens. Methods GHESKIO (Haitian Study Group for Kaposi's sarcoma and Opportunistic Infections) clinic attendees 18 years of age or older were invited to participate. Venipuncture blood specimens were used for reference testing: for HIV, Murex HIV-1.2.0 (Dia-Sorin S.p. A.) or Determine HIV-1/2 (Alere Inc). Positive results were confirmed with the HIV(1+2) Rapid Test Strip (KHB Shanghai Kehua Bioengineering Co. Ltd). For Treponema pallidum (Tp) antibody comparison, Treponema Pallidum Hemagglutination Assay (TPHA) (Human Gesellschaft fur Biochemica und Diagnostica mbH) was used. For 21 TPHA indeterminate results, specimens were retested using a Tp enzyme-linked immunosorbent assay test (ELISA) (Architect Syphilis Tp, Abbott Laboratories). Sensitivity and specificity were calculated and the exact binomial method was used to determine 95% confidence intervals (CI). Results Of 298 study participants, 61 (20.5%) were male. Of 237 females, 49 (20.7%) were pregnant. For the HIV component, sensitivity and specificity were 99.2% (95% CI: 95.8%, 100%) and 97.0% (95% CI: 93.2%, 99.0%), respectively. All 21 TPHA indeterminate results were Tp ELISA reactive. For the Tp component, sensitivity and specificity were 96.5% (95% CI: 91.2%, 99.0%) and 90.8% (95% CI: 85.7%, 94.6%), respectively. In pregnant women, the HIV component sensitivity and specificity were 93.3% (95% CI: 68.0%, 99.8%) and 94.1% (95% CI: 80.3%, 99.3%), respectively; and for the Tp component were 100% (95% CI: 81.5%, 100%) and 96.8% (83.3%, 99.9%), respectively. Conclusion The HIV antibody component of the Duo test shows excellent performance. The Treponema pallidum antibody component showed high sensitivity, and slightly lower specificity. Amongst pregnant women the test performed very well. Abstracts Sex Transm Infect 2015;91(Suppl 2):A1-A258 A129