A rapid economical multiplex PCR-RFLP method for molecular detection and genotyping of Giardia duodenalis clinical isolates

IF 2.1 4区医学 Q3 INFECTIOUS DISEASES

Diagnostic microbiology and infectious disease Pub Date : 2024-10-03 DOI:10.1016/j.diagmicrobio.2024.116548

Ajanta Ghosal , Sanjib K. Sardar , Tapas Haldar , Akash Prasad , Koushik Das , Seiki Kobayashi , Yumiko Saito-Nakano , Shanta Dutta , Tomoyoshi Nozaki , Sandipan Ganguly

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Abstract

Giardia duodenalis is a common cause of diarrheal illness in regions with limited resources. The demand for rapid and cost-effective detection and genotyping methods in large-scale epidemiological studies and clinical diagnostics is imperative. Hence, we developed a multiplex PCR-RFLP technique targeting the tpi gene of G. duodenalis. The assay successfully screened G. duodenalis positive clinical samples (6.33 %; 36/565). It was also able to categorize the isolates into assemblages A (41.66 %; 13/36) and B (58.33 %; 23/36), as well as into subassemblages: AI (13.8 %; 5/36), AII (27.77 %; 10/36), BIII (36.11 %; 15/36) and BIV (22.22 %; 8/36). High diagnostic sensitivity (94.2 %), specificity (100 %) and accuracy (97.1 %) of the PCR assay were obtained, indicating its reliability for diagnosing giardiasis. Notably, the assay demonstrated close concordance with microscopy (κ=0.85) and reference PCR (κ=0.98) results. The optimized method offers a cost-effective and rapid approach for G. duodenalis detection and genotyping, convenient for epidemiological studies and clinical diagnostics.

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用于对十二指肠贾第虫临床分离株进行分子检测和基因分型的快速、经济的多重 PCR-RFLP 方法。

在资源有限的地区，十二指肠贾第虫是腹泻病的常见病因。在大规模流行病学研究和临床诊断中，对快速、经济有效的检测和基因分型方法的需求势在必行。因此，我们开发了一种针对十二指肠杆菌 tpi 基因的多重 PCR-RFLP 技术。该检测方法成功筛查了十二指肠杆菌阳性临床样本（6.33%；36/565）。它还能将分离物分为 A 组合（41.66%；13/36）和 B 组合（58.33%；23/36），以及亚组合：AI（13.8%；5/36）、AII（27.77%；10/36）、BIII（36.11%；15/36）和 BIV（22.22%；8/36）。PCR 检测法的诊断灵敏度（94.2%）、特异度（100%）和准确度（97.1%）都很高，这表明它在诊断贾第虫病方面非常可靠。值得注意的是，该检测方法与显微镜检测（κ=0.85）和参考 PCR 检测（κ=0.98）结果非常吻合。优化后的方法为十二指肠杆菌的检测和基因分型提供了一种经济、快速的方法，为流行病学研究和临床诊断提供了便利。

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来源期刊

Diagnostic microbiology and infectious disease 医学-传染病学

CiteScore

5.30

自引率

3.40%

发文量

149

审稿时长

56 days

期刊介绍： Diagnostic Microbiology and Infectious Disease keeps you informed of the latest developments in clinical microbiology and the diagnosis and treatment of infectious diseases. Packed with rigorously peer-reviewed articles and studies in bacteriology, immunology, immunoserology, infectious diseases, mycology, parasitology, and virology, the journal examines new procedures, unusual cases, controversial issues, and important new literature. Diagnostic Microbiology and Infectious Disease distinguished independent editorial board, consisting of experts from many medical specialties, ensures you extensive and authoritative coverage.