Evaluation of rapid method for detection of cytomegalovirus in clincal specimens using polymerase chain reaction DNA amplification

Human cytomegalovirus (HCMV) infection is the major cause of illness and death in immunocompromised patients. HCMV is the most common cause of congenital viral infection in humans. A polymerase chain reaction (PCR) method was developed for the rapid detection of CMV in urine. Several parameters of the PCR procedure were optimized to reduce time and improve sensitivity. By eliminating the extraction of DNA from clinical specimens, reducing the number of amplification cycles, utilization of the hot start PCR procedure and direct detection of PCR product by ethidium bromide fluorescence staining, a procedure was developed which could be performed in less than 3 hours. Comparison studies using cell culture and direct detection of CMV by PCR on urine specimens were performed. Sensitivity was further examined to determine if inhibitors of the PCR reaction were present in urine