Development of qPCR Detection Kit for Langya Henipavirus L Gene

Abstract

Langya Henipavirus (LayV) is a new zoonotic Henipavirus reported in 2022. Patients infected by LayV show common cold symptoms, such as fever, fatigue, and cough. Non-fatal cases associated with this virus have been reported in Henan and Shandong provinces in China. Should human-to-human transmission occur, it has the potential to be a pandemic like SARS-CoV-2. LayV is an enveloped negative-sense RNA virus. Its polymerase (encoded by the L gene) is considered a conserved target shared within Henipavirus viruses, allowing for qPCR detection, in which primers play a pivotal role. For the development of a diagnostic kit, we designed three primer sets targeting the L gene and evaluated their detection of plasmids carrying part of the L gene. Primer sets 1 and 2 were found to be suitable for further sensitivity, specificity, and efficiency testing. Both primer sets were able to amplify PCR products in the presence of human saliva, and both the melt curve analysis and gel electrophoresis showed distinct peaks and bands, supporting the specificity of the qPCR primers. Based on standard curve analysis, both primers detected as low as approximately 1000 copies of the L gene. Given the specificity and robustness to work in the presence of saliva, the primers can be used for pandemic-preparedness in LayV detection.