VOLUME 13 (Supplement)

Development of a closed-tube QUASR-LAMP assay for the detection of human adenoviruses

Human adenoviruses (HAdVs) are double-stranded DNA viruses capable of causing a wide array of diseases in humans. They are ubiquitous in nature and may cause life-threatening opportunistic infections in immunocompromised individuals. Diagnosis is traditionally by cell culture or antigen detection methods. Development of rapid diagnostic kits for the detection of enteric adenoviruses can help to rule out other underlying causes of gastroenteritis in patients. Also, these kits may be used to detect HAdVs in environmental samples. This present study presented the development of a quenching of unincorporated amplification signal reporters–loop-mediated isothermal amplification (QUASR–LAMP) assay to detect HAdVs for clinical and environmental applications. The optimized assay can be completed in 50 min at 63-64 ºC. The minimum detection limit of the QUASR–LAMP is 50 copies/µL while the polymerase chain reaction (PCR) assay was 100 copies/µL. Thirteen (13) human fecal samples and 6 water samples were used to determine the measures of accuracy. The sensitivity of the assay was 85.71%, while the specificity was 92.86%. The positive predictive value (PPV) was 82.76% while the negative predictive value (NPV) was 94.20% and an accuracy of 90.82%. The assay is also specific to HAdV DNA. The QUASR–LAMP assay can potentially be modified and applied as a simple diagnostic tool to detect infections in humans and to determine water quality for risk assessment especially in resource-limited areas.