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BACKGROUND: Nested nucleic acid amplification tests are often thought too sensitive or prone to generatingfalse positive results for routine use. The current study investigated the specificity and clinicalutility of a routine multiplex nested assay for mucosal herpetic infections. METHODS: Ninety patients, categorised into those clinically diagnosed to (a) have and (b) not haveherpetic infection, were enrolled. Swabs from oral and ano-genital sites were assayed by thenested assay and culture and the results assessed against clinical evaluation for diagnosingherpetic infections; cell content was also recorded. RESULTS: Twenty-six and 64 patients were thought to (a) have and (b) not have mucosal herpeticinfection. Taking the clinical evaluation as indicating the presence of herpetic infection, thenested polymerase chain reaction and culture had respective sensitivities of 19/26 (73%) and12/26 (46%) (Χ2 p = 0.02). There was no significant difference in specificities between nPCR62/64 (97%) and culture 63/64 (98%) (Χ2 p = 1.0). Cell content was important for viraldetection by nPCR (Χ2 p = 0.07) but not culture. Nesting was found necessary for sensitivity anddid not reduce specificity. Assay under-performance appeared related to sub-optimal cellcontent (20%) but may have reflected clinical over-diagnosis. The results suggest the need forvalidating specimen cell quality. CONCLUSIONS: This study questions the value of routine laboratory confirmation of mucosal herpetic infection. The adoption of a more discriminatory usage of laboratory diagnostic facilities for genital herpetic infection, taking account of cell content, and restricting it to those cases where it actually affects patient management, may be warranted.

bmc infectious diseases

Clinical utility of a nested nucleic acid amplification format in comparison to viral culture for the diagnosis of mucosal herpes simplex infection in a genitourinary medicine setting