Development and evaluation of a novel in-house single-tube nested real-time polymerase chain reaction assay with FilmArray respiratory panel for detection of adenovirus

Abstract

FilmArray is a rapid diagnostic technology which can simultaneously detect multiple pathogens in a single run. It is especially useful in the diagnosis of respiratory diseases because most of the causative pathogens cause similar symptoms. While FilmArray has numerous undisputable advantages in the detection of respiratory pathogens, one of the major concerns is that it has poor sensitivity for adenovirus detection. Reports are consistently showing that the FilmArray Respiratory Panel has only around 50% to 60% sensitivity for the detection of adenoviruses. FilmArray Respiratory Panel’s poor sensitivity for adenovirus detection is of particular concern for immunocompromised patients because they are at higher risk of developing severe infection. Therefore, a complementary test would be needed to overcome these limitations of FilmArray. In this study, a novel in-house single-tube nested real-time polymerase chain reaction assay was developed and we evaluated its performance with that of FilmArray Respiratory Panel for adenovirus detection. Improved performance for detection of adenoviruses has been shown for the in-house single-tube nested real-time polymerase chain reaction assay. Eight additional samples that were tested negative by FilmArray Respiratory Panel were tested positive by the single-tube nested real-time polymerase chain reaction assay, with low viral loads of 3.2 to 1.8 x 〖10〗^4 DNA copies/reaction. The use of this single-tube nested real-time polymerase chain reaction assay should be considered in patients with clinical suspicion of adenovirus but a negative FilmArray Respiratory Panel result.