A detection method for the rash and fever illness ‐associated viruses using multiplex RT‐PCR

ABSTRACT To detect various viral genes in the patients with rash and fever illness (RFI), a new multiplex (reverse transcription) polymerase chain reaction [(RT)‐PCR] method was constructed. New primer sets were designed for the detection of herpes simplex viruses 1 and 2 (HSV1 and 2), and Epstein‐Barr virus (EBV). The newly designed and previously reported primer sets were used to detect 13 types of RFI‐associated viruses by multiplex (RT)‐PCR assay systems. Moreover, to eliminate non‐specific PCR products, a double‐stranded specific DNase was used for the digestion of double‐stranded DNA derived from the templates in clinical specimens. RFI‐associated viruses were detected in 77.0% of the patients (97/126 cases) by the presented method. Multiple viruses were identified in 27.8% of the described cases (35/126 cases). The compatibilities between detected viruses and clinical diagnosis were found in 32.5% of the patients (41/126 cases). Sensitivity limits of these viruses were estimated to be 101–103 copies/assay. Furthermore, non‐specific PCR products were eliminated by a double‐stranded specific DNase with no influence on the sensitivity. The results suggested that this method can detect various RFI‐associated viruses with high sensitivity and specificity in clinical specimens.
Source: Microbiology and Immunology - Category: Microbiology Authors: Tags: Original Article Source Type: research