A-093 Molecular Detection of Rabies Using the LN34 Pan-Lyssavirus Real-Time RT-PCR Assay

IF 8.3 2区医学 Q1 MEDICAL LABORATORY TECHNOLOGY

Clinical chemistry Pub Date : 2026-10-04 DOI:10.1093/clinchem/hvag086.069

Bryan Nguyen, Mariam Zhowandai, Mihyun Kim, Megan Crumpler, Radhika Patel

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引用次数: 0

Abstract

Background Orange County Public Health Laboratory (OCPHL) routinely performs diagnostic rabies virus testing on animal brain material using the CDC Protocol for Postmortem Diagnosis of Rabies in Animals by Direct Fluorescent Antibody (DFA). The objective of this study was to validate the CDC LN34 qRT-PCR assay, which detects rabies virus RNA, using the Applied BiosystemsTM ABI 7500 Fast Dx. Implementing the LN34 assay included the validation and optimization of a safe Rabies virus RNA extraction method with the Qiagen TissueLyser II and the Qiagen EZ1 Advanced XL instrument. Methods Forty animal brain tissue specimens, previously tested for rabies virus by DFA, were tested on the Fusion LN34 assay. Results were compared with DFA as the gold standard. The CDC LN34 qRT-PCR assay was validated using the EZ1 instrument for viral RNA extraction. An inactivation study was performed to test the viability of rabies virus in Hologic Specimen Transport Media using cell cultures of serial ten-fold dilutions. The inactivation study was performed by Wadsworth Center’s Griffin Laboratory at New York State Department of Health. Results The validation of the LN34 had a 100% accuracy and 100% precision with the qualitative data in comparison with rabies DFA. Inactivation study in progress. Conclusion With the validation of the LN34 assay, rabies diagnostics can become more sensitive, specific, and economical. Currently, the LN34 assay serves to be an additional confirmatory test following the gold standard, direct fluorescence antibody (DFA) microscopy.

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LN34泛溶血病毒实时RT-PCR检测狂犬病毒A-093

橙县公共卫生实验室（OCPHL）根据美国疾病控制与预防中心（CDC）动物死后狂犬病直接荧光抗体（DFA）诊断方案，对动物脑材料进行常规狂犬病毒诊断检测。本研究的目的是验证CDC LN34 qRT-PCR检测狂犬病病毒RNA，使用Applied BiosystemsTM ABI 7500 Fast Dx。实施LN34检测包括使用Qiagen TissueLyser II和Qiagen EZ1 Advanced XL仪器验证和优化一种安全的狂犬病病毒RNA提取方法。方法采用融合LN34法对40例动物脑组织标本进行狂犬病毒检测。结果与DFA作为金标准进行了比较。采用EZ1病毒RNA提取仪验证CDC LN34 qRT-PCR检测方法。采用连续十倍稀释的细胞培养，进行了一项灭活研究，以测试狂犬病毒在Hologic标本运输介质中的生存能力。灭活研究是由纽约州卫生部沃兹沃斯中心的格里芬实验室进行的。结果与狂犬病DFA的定性数据比较，LN34的准确度为100%，精密度为100%。灭活研究正在进行中。结论通过对LN34检测方法的验证，可以提高狂犬病诊断的灵敏性、特异性和经济性。目前，LN34检测是继金标准——直接荧光抗体（DFA）显微镜之后的一项额外的验证性检测。

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

Clinical chemistry 医学-医学实验技术

CiteScore

11.30

自引率

4.30%

发文量

212

审稿时长

1.7 months

期刊介绍： Clinical Chemistry is a peer-reviewed scientific journal that is the premier publication for the science and practice of clinical laboratory medicine. It was established in 1955 and is associated with the Association for Diagnostics & Laboratory Medicine (ADLM). The journal focuses on laboratory diagnosis and management of patients, and has expanded to include other clinical laboratory disciplines such as genomics, hematology, microbiology, and toxicology. It also publishes articles relevant to clinical specialties including cardiology, endocrinology, gastroenterology, genetics, immunology, infectious diseases, maternal-fetal medicine, neurology, nutrition, oncology, and pediatrics. In addition to original research, editorials, and reviews, Clinical Chemistry features recurring sections such as clinical case studies, perspectives, podcasts, and Q&A articles. It has the highest impact factor among journals of clinical chemistry, laboratory medicine, pathology, analytical chemistry, transfusion medicine, and clinical microbiology. The journal is indexed in databases such as MEDLINE and Web of Science.