Evaluation of a semi-automated method for detection of cytomegalovirus DNA in multiple sample types.

Cytomegalovirus (CMV) is the most common congenital infection and an important cause of childhood disability. Newborn CMV screening and studies of mother-to-child transmission are increasingly common, involving viral detection using various sample types. Here, we describe the analytical evaluation of the AltoStar® CMV PCR Kit 1.5, a commercial diagnostic platform for detection of CMV DNA in diverse sample types relevant for congenital CMV infection. Oral swabs, saliva, dried urine spots (DUS) and dried blood filter paper spots (DBS), umbilical cord blood, amniotic fluid, placental tissue, vaginal swabs, and breast milk from known CMV-negative donors were spiked with known concentrations of viral isolates. Linearity of detection in fluid samples ranged from 0.96 to 1.00. Limits of detection in fluid samples ranged from 2.22 to 2.83 log IU/ml. CMV PCR on dried urine spot samples had greater sensitivity when using off-instrument nucleic acid purification with Quanta Extracta DBS buffer (QuantaBio). This study provides a cross-matrix performance framework for CMV molecular testing in congenital infection research and newborn screening contexts. The AltoStar® CMV PCR platform demonstrated robust performance across clinically relevant specimen types and scalable throughput capacity, supporting its suitability for large-scale epidemiologic and screening applications.