Development and evaluation of a quantitative assay detecting cytomegalovirus transcripts for preemptive therapy in...

Successful preemptive therapy for cytomegalovirus (CMV) infection after hematopoietic stem cell transplantation (HSCT) depends on the availability of a rapid and sensitive assay to guide early treatment. Currently, the antigenemia assay and the quantitative real-time PCR (qPCR) assay are widely used for this purpose, but they have distinctive concerns. This study aimed to develop and evaluate a novel CMV diagnostic test based on transcription-reverse transcription concerted reaction (TRC), an RNA-detecting technology. The CMV-TRC assay detected CMV β2.7 transcripts within 10 min over a five-log range. Among a total of 219 samples obtained from 24 allogeneic HSCT recipients, samples detected as positive by the CMV-TRC assay showed a relatively strong correlation with those detected as positive by the qPCR assay and the antigenemia assay. The CMV-TRC assay showed higher sensitivity (77.7%) than the antigenemia assay (68.1%) and detected the first and recurrent episodes of active CMV infection in HSCT patients significantly earlier than the antigenemia assay (P < 0.001). Although the CMV-TRC assay (87.8%) showed low sensitivity compared to the qPCR assay (96.3%), the performance of the CMV-TRC assay was equivalent to that of the qPCR assay in detecting the appearance of active CMV infection episodes (P < 0.092) or rather superior in detecting the clearance of episodes (P < 0.001). The CMV-TRC assay with several advantages may be useful for guiding preemptive anti-CMV therapy in HSCT recipients.