Version 1
: Received: 16 January 2024 / Approved: 17 January 2024 / Online: 17 January 2024 (09:13:56 CET)
Version 2
: Received: 20 January 2024 / Approved: 22 January 2024 / Online: 22 January 2024 (10:42:23 CET)
How to cite:
Yuan, K.; Zhu, H.; Jiang, T.; Xu, M.; Du, S.; Gan, S.K. Development of qPCR Detection Kit for Langya Henipavirus L Gene. Preprints2024, 2024011298. https://doi.org/10.20944/preprints202401.1298.v2
Yuan, K.; Zhu, H.; Jiang, T.; Xu, M.; Du, S.; Gan, S.K. Development of qPCR Detection Kit for Langya Henipavirus L Gene. Preprints 2024, 2024011298. https://doi.org/10.20944/preprints202401.1298.v2
Yuan, K.; Zhu, H.; Jiang, T.; Xu, M.; Du, S.; Gan, S.K. Development of qPCR Detection Kit for Langya Henipavirus L Gene. Preprints2024, 2024011298. https://doi.org/10.20944/preprints202401.1298.v2
APA Style
Yuan, K., Zhu, H., Jiang, T., Xu, M., Du, S., & Gan, S.K. (2024). Development of qPCR Detection Kit for Langya Henipavirus L Gene. Preprints. https://doi.org/10.20944/preprints202401.1298.v2
Chicago/Turabian Style
Yuan, K., Shihan Du and Samuel Ken-En Gan. 2024 "Development of qPCR Detection Kit for Langya Henipavirus L Gene" Preprints. https://doi.org/10.20944/preprints202401.1298.v2
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.
Keywords
Langya Henipavirus; primer design; qPCR; detection kit
Subject
Biology and Life Sciences, Virology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
