反转录重组酶辅助扩增与侧流试纸和实时荧光相结合的戊型肝炎病毒快速视觉检测。

IF 6.1 2区 医学 Q1 MICROBIOLOGY Journal of Clinical Microbiology Pub Date : 2025-02-19 Epub Date: 2025-01-16 DOI:10.1128/jcm.01064-24
Bingyan Wei, Wenlong Wang, Zixuan Guo, Wenjiao Yin, Minheng Cheng, Yifei Yang, Yuewei Tian, Yaxin Sun, Tianlong Liu, Yanxin Hu, Ruiping She, Jijing Tian
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引用次数: 0

摘要

戊型肝炎病毒(HEV)是一种全球流行的人畜共患病原体,主要通过粪-口途径传播,例如食用未煮熟或受污染的猪肉。估计每年在人群中,戊型肝炎感染导致330万病毒性肝炎症状病例和7万人死亡。因此,一种快速准确的方法检测血清或粪便样本中的戊肝病毒是必不可少的。在这项研究中,我们旨在开发和评估两种快速简便的检测HEV RNA的方法:逆转录重组酶辅助横向流动试尺扩增(RT-RAA-LFD)和定量实时逆转录重组酶辅助扩增(qRT-RAA)。我们优化了反应条件,并评估了它们的敏感性和特异性。RT-RAA-LFD在39°C下15分钟内完成反应,检测限(LOD)为247拷贝/μL,达到95%。qRT-RAA测定在42°C下20分钟内完成,LOD为25拷贝/μL,为95%。两种检测方法均显示与其他猪病原体无交叉反应性,且具有很强的特异性。RT-RAA-LFD法检测245份猪胆汁和粪便,kappa值为0.943 (P < 0.001),与定量反转录PCR符合率为97.14%(238/245)。同样,qRT-RAA法kappa值为0.976 (P < 0.001),符合率为98.78%(242/245)。总之,这两种RT-RAA检测方法有望成为兽医诊所广泛和有效筛查猪戊型肝炎的有效诊断工具。重要性:戊型肝炎病毒(HEV)是一种全球广泛传播的人畜共患病原体,对公共卫生构成重大风险。猪是人畜共患戊型肝炎的主要天然宿主。本研究介绍了一种快速、精确的猪HEV RNA检测方法,显示了其作为兽医诊所全面、高效筛选猪HEV的有效诊断工具的潜力。
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Rapid visual detection of hepatitis E virus combining reverse transcription recombinase-aided amplification with lateral flow dipstick and real-time fluorescence.

Hepatitis E virus (HEV) is a globally prevalent zoonotic pathogen that is primarily spread through the fecal-oral route, such as by consuming undercooked or contaminated pork. HEV infection leads to an estimated 3.3 million symptomatic cases of viral hepatitis and 70,000 deaths in human populations each year. Therefore, a rapid and accurate method for detecting HEV in serum or stool samples is essential. In this study, we aimed to develop and evaluate two methods for the rapid and convenient detection of HEV RNA: reverse transcription recombinase-aided amplification with lateral flow dipstick (RT-RAA-LFD) and quantitative real-time reverse transcription recombinase-aided amplification (qRT-RAA). We optimized the reaction conditions and assessed their sensitivity and specificity. The RT-RAA-LFD assay completed its reaction at 39°C within 15 minutes, achieving a 95% limit of detection (LOD) of 247 copies/μL. The qRT-RAA assay, completed at 42°C within 20 minutes, had a 95% LOD of 25 copies/μL. Both assays demonstrated no cross-reactivity with other porcine pathogens and exhibited strong specificity. In testing 245 porcine bile and fecal samples, the RT-RAA-LFD assay showed a kappa value of 0.943 (P < 0.001) with a 97.14% (238/245) coincidence rate compared with quantitative reverse transcription PCR. Similarly, the qRT-RAA assay achieved a kappa value of 0.976 (P < 0.001) with a 98.78% (242/245) coincidence rate. In conclusion, these two RT-RAA assays show promising potential as effective diagnostic tools for broad and efficient screening of swine HEV in veterinary clinics.

Importance: Hepatitis E virus (HEV) is a globally widespread zoonotic pathogen that poses a significant public health risk. Swine serve as the primary natural host for zoonotic HEV. This study introduces a rapid and precise method for detecting swine HEV RNA, showcasing its potential as an effective diagnostic tool for comprehensive and efficient screening of swine HEV in veterinary clinics.

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来源期刊
Journal of Clinical Microbiology
Journal of Clinical Microbiology 医学-微生物学
CiteScore
17.10
自引率
4.30%
发文量
347
审稿时长
3 months
期刊介绍: The Journal of Clinical Microbiology® disseminates the latest research concerning the laboratory diagnosis of human and animal infections, along with the laboratory's role in epidemiology and the management of infectious diseases.
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