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磁珠-金纳米颗粒-PCR扩增免疫分析法检测胸膜结核病例中的MPT-64蛋白

Detection of MPT-64 protein in pleural tuberculosis cases by magnetic bead-gold nanoparticle-PCR amplified immunoassay.

作者信息

Soni Aishwarya, Nehra Kiran, Dahiya Bhawna, Rais Anam, Prasad Tulika, Gahlaut Anjum, Raj Vikas, Sheoran Reetu, Parmar Aparna, Mehta Promod K

机构信息

Microbiology Department, Faculty of Allied Health Sciences (FAHS), Shree Guru Gobind Singh Tricentenary (SGT) University, Gurugram, India.

Department of Biotechnology, Deenbandhu Chhotu Ram University of Science and Technology (DCRUST), Sonipat, India.

出版信息

Future Microbiol. 2025 Feb;20(2):107-115. doi: 10.1080/17460913.2024.2432179. Epub 2024 Nov 29.

Abstract

AIM

Diagnosis of pleural tuberculosis (TB) is challenging; thus, an efficient method is urgently needed.

METHODS

We developed a magnetic-bead-gold nanoparticle-PCR amplified immunoassay (MB-AuNP-I-PCR, liquid system) to detect the MPT-64 protein in pleural TB patients. AuNPs functionalized with detection antibodies/oligonucleotides were characterized by UV-vis spectroscopy, Transmission/Scanning electron microscopy, Fourier-transform infrared spectrometer, ELISA, and PCR, whereas MBs conjugated with detection antibodies were validated by magneto-ELISA/UV-vis spectroscopy.

RESULTS

We utilized the MB-AuNP-I-PCR for MPT-64 detection in 99 clinical specimens which displayed 85.2% sensitivity and 97.8% specificity to diagnose pleural TB cases. Markedly, the sensitivity achieved by MB-AuNP-I-PCR was noticeably higher ( < 0.01) than magneto-ELISA and GeneXpert.

CONCLUSION

This is a preliminary report to diagnose pleural TB cases by MB-AuNP-I-PCR with promising results that require further corroboration in a higher number of specimens.

摘要

目的

胸膜结核(TB)的诊断具有挑战性,因此迫切需要一种有效的方法。

方法

我们开发了一种磁珠-金纳米颗粒-PCR扩增免疫测定法(MB-AuNP-I-PCR,液体系统)来检测胸膜结核患者的MPT-64蛋白。用检测抗体/寡核苷酸功能化的金纳米颗粒通过紫外-可见光谱、透射/扫描电子显微镜、傅里叶变换红外光谱仪、酶联免疫吸附测定和PCR进行表征,而与检测抗体偶联的磁珠则通过磁酶联免疫吸附测定/紫外-可见光谱进行验证。

结果

我们利用MB-AuNP-I-PCR对99份临床标本进行MPT-64检测,在诊断胸膜结核病例时显示出85.2%的灵敏度和97.8%的特异性。显著的是,MB-AuNP-I-PCR所达到的灵敏度明显高于磁酶联免疫吸附测定和GeneXpert(<0.01)。

结论

这是一篇关于用MB-AuNP-I-PCR诊断胸膜结核病例的初步报告,结果令人鼓舞,但需要在更多标本中进一步证实。

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