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

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Analytical and Bioanalytical Chemistry Pub Date : 2024-12-16 DOI:10.1007/s00216-024-05690-2

Jiayi Yang, Mingwei Liu, Huijie Li, Yunhua Gao, Lianhua Dong

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引用次数: 0

Abstract

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.

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开发轮状病毒 G3P8 和 G9P8 的 RT-dPCR 方法和参考材料。

轮状病毒（RV）是 5 岁以下儿童急性肠胃炎的主要致病病原体，每年导致全球 20 万婴儿死亡。尽管已开发出疫苗，但它仍加重了世界各地的医疗负担。在此，我们开发了反转录数字 PCR（RT-dPCR）方法，用于精确和绝对量化轮状病毒 G3P8 和 G9P8 的核酸。假病毒参考材料（RM）包含编码 VP4 和 VP7 的 RNA 片段。不确定度扩大后的分配值分别为 (2432 ± 510) copies/μL 和 (3406 ± 613) copies/μL。采用 RM 和 RT-dPCR 方法对各种数字平台进行了验证，发现平台 III 的性能不足，有可能导致 "假阴性 "结果。RT-dPCR 技术和假病毒 RM 的应用在诊断 RV 引起的疾病方面具有优势，从而提高了急性肠胃炎患儿的存活率。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Analytical and Bioanalytical Chemistry 化学-分析化学

CiteScore

8.00

自引率

4.70%

发文量

638

审稿时长

2.1 months

期刊介绍： Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.