开发新型基因芯片和限制性片段长度多态性(RFLP)方法,用于肉汤培养液中结核分枝杆菌复合体的快速检测。

IF 4.5 2区 医学 Q2 IMMUNOLOGY Journal of Microbiology Immunology and Infection Pub Date : 2024-09-21 DOI:10.1016/j.jmii.2024.09.003
Wen-Hung Wang, Chun-Yu Lin, Shu-Huei Jain, Po-Liang Lu, Yen-Hsu Chen
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引用次数: 0

摘要

背景:结核病(TB)是一个重大的全球公共卫生问题。及时准确的结核病诊断对于开始适当的治疗和防止疾病蔓延至关重要。目前的诊断技术要么缓慢,要么昂贵。本研究旨在利用基因芯片和对分枝杆菌生长指示管(MGIT)标本的限制性片段长度多态性(RFLP)分析,创建并评估一种新型、快速、高度可靠且经济高效的结核病检测方法:我们在 2000 份 MGIT 培养阳性标本中评估了针对结核分枝杆菌 16S rRNA 基因的新型基因芯片和 RFLP 方法的有效性。RFLP 分析确定了结核分枝杆菌复合体(MTBC)基因组中的 AfeI 限制位点。通过广泛的测序和 Cobas TaqMan PCR 分析,并查阅患者资料,对差异进行了调查:基因芯片法的灵敏度为 99.27%,特异性为 98.35%;RFLP 法的灵敏度为 98.18%,特异性为 99.31%。两个分离物的假阴性是由于 AfeI 位点发生了突变。此外,有五个病例在培养物中生长出非结核分枝杆菌时显示出 MTBC 的存在:结论:我们的新型基因芯片和 RFLP 方法可在 MGIT 标本中快速、高可靠、低成本地检测结核杆菌。基因芯片和 RFLP 方法都适用于资源有限的环境,具有经济优势。这些方法在改善临床结核病诊断方面具有巨大潜力。
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Development of the novel gene chip and restriction fragment length polymorphism (RFLP) methods for rapid detection of Mycobacterium tuberculosis complex in broth culture.

Background: Tuberculosis (TB) is a major global public health issue. Prompt and accurate TB diagnosis is crucial for starting appropriate treatments and preventing the disease's spread. Current diagnostic techniques are either slow or expensive. This study aimed to create and evaluate a new, fast, highly reliable, and cost-effective TB detection method using a gene chip and Restriction Fragment Length Polymorphism (RFLP) analysis on Mycobacteria Growth Indicator Tubes (MGIT) specimens.

Methods: We assessed the effectiveness of a novel gene chip and RFLP methods targeting the 16S rRNA gene of Mycobacterium tuberculosis in 2000 MGIT culture-positive specimens. RFLP analysis identified the AfeI restriction site within the M. tuberculosis complex (MTBC) genome. Discrepancies were investigated through extensive sequencing and Cobas TaqMan PCR analysis, along with reviewing patient profiles.

Results: Both methods showed high efficacy in detecting MTBC in broth cultures, with the gene chip method achieving a sensitivity of 99.27 %, specificity of 98.35 %, and the RFLP method showing a sensitivity of 98.18 %, specificity of 99.31 %. False negatives in two isolates were due to a mutation in the AfeI site. Additionally, five cases showed MTBC presence when nontuberculous Mycobacterium species grew in cultures.

Conclusion: Our novel gene chip and RFLP methods are effective for rapid highly-reliable and cost-effective M. tuberculosis detection in MGIT specimens. Both gene chip and RFLP methods are suitable for resource-limited settings, offering an economical advantage. These methods have significant potential to improve clinical TB diagnosis.

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来源期刊
Journal of Microbiology Immunology and Infection
Journal of Microbiology Immunology and Infection IMMUNOLOGY-INFECTIOUS DISEASES
CiteScore
15.90
自引率
5.40%
发文量
159
审稿时长
67 days
期刊介绍: Journal of Microbiology Immunology and Infection is an open access journal, committed to disseminating information on the latest trends and advances in microbiology, immunology, infectious diseases and parasitology. Article types considered include perspectives, review articles, original articles, brief reports and correspondence. With the aim of promoting effective and accurate scientific information, an expert panel of referees constitutes the backbone of the peer-review process in evaluating the quality and content of manuscripts submitted for publication.
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