Detection of adenoviruses in feces of infants with diarrhea by real-time PCR

Abstract

A Real-time PCR method was established to study the infection of adenovirus (Ad) in infants with sporadic diarrhea in Wenzhou. According to hexon gene of adenovirus, one prime pair was designed as universal primes and applied to detect adenovirus DNA by Real-time PCR. It was also compared with immunochromatographic assay. 157 fecal specimens from diarrhea infants were tested while positive specimens were sequenced and identified by isolate culture and restriction endonucleases. A rapid and specific Real-time PCR assay for detection adenovirus was set up. The positive rates of adenovirus in fecal specimens by immunochromatographic assay and Real-time PCR were 1.91% (3/157) and 3.18% (5/157), respectively. Out of the 154 specimens with negative result from immunochromatographic assay, 2 showed positive by Real-time PCR. 5 positive specimens, identified by Real-time PCR, were sequenced as Ad3 (3/157, 1.91% ) and Ad7 (2/157, 1.27%). 2 of the 5 positive specimens were proved to be Ad3 by cell culture and restriction endonucleases. Real-time PCR combined with sequence analysis seemed more sensitive and specific so could be used for identifying types of adenovirus in clinical specimens. Ad3 and Ad7 were important pathogens which caused infant sporadic diarrhea in Wenzhou during February and April in 2008.