Development of RT-dPCR method and reference material for rotavirus G3P8 and G9P8.

Autor: Yang J; Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100029, China. yangjy2018@nim.ac.cn., Liu M; Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100029, China., Li H; Shenzhen Institute for Technology Innovation, NIM, Shenzhen, 518132, China., Gao Y; Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100029, China., Dong L; Center for Advanced Measurement Science, National Institute of Metrology, Beijing, 100029, China.
Jazyk: angličtina
Zdroj: Analytical and bioanalytical chemistry [Anal Bioanal Chem] 2024 Dec 16. Date of Electronic Publication: 2024 Dec 16.
DOI: 10.1007/s00216-024-05690-2
Abstrakt: The rotavirus (RV) is the predominant causative pathogen associated with acute gastroenteritis in children aged below 5 years, leading to an annual mortality rate of 200,000 infants globally. Despite the development of a vaccine, it exacerbates the medical burden around the world. Here, we have developed reverse transcription digital PCR (RT-dPCR) methods for precise and absolute quantification of nucleic acid in rotavirus G3P8 and G9P8. The pseudovirus reference material (RM) contained the RNA fragment encoding VP4 and VP7. The assigned values with expanded uncertainty were determined as (2432 ± 510) copies/μL and (3406 ± 613) copies/μL. The RM and RT-dPCR methods were employed to validate various digital platforms, revealing the inadequate performance of platform III, which could potentially result in "false-negative" outcomes. The application of RT-dPCR techniques and pseudovirus RM confers advantages in the diagnosis of RV-induced diseases, thereby enhancing the survival rate of young children suffering from acute gastroenteritis.
Competing Interests: Declarations. Competing interests: The authors declare no competing interests.
(© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)
Databáze: MEDLINE
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