基于纳米的实时免疫聚合酶链反应对肺外结核病例中分枝杆菌蛋白的定量分析
Quantification of mycobacterial proteins in extrapulmonary tuberculosis cases by nano-based real-time immuno-PCR.
作者信息
Dahiya Bhawna, Prasad Tulika, Rais Anam, Sheoran Abhishek, Kamra Ekta, Mor Preeti, Soni Aishwarya, Sharma Suman, Mehta Promod K
机构信息
Centre for Biotechnology, Maharshi Dayanand University, Rohtak, 124001, Haryana, India.
Special Centre for Nano Science & Advanced Instrumentation Research Facility, Jawaharlal Nehru University, New Delhi, 110067, India.
出版信息
Future Microbiol. 2023 Aug;18:771-783. doi: 10.2217/fmb-2023-0054. Epub 2023 Aug 4.
Diagnosis of extrapulmonary tuberculosis (EPTB) is difficult, and a rapid and dependable diagnostic test is urgently needed. A nano-based assay, SYBR Green magnetic bead-coupled gold nanoparticle-based real-time immuno-polymerase chain reaction (MB-AuNP-RT-I-PCR) was studied for the quantitative detection of MPT-64+CFP-10 proteins in clinically suspected EPTB patients. A wide range (270 fg/ml-9.9 ng/ml) of MPT-64+CFP-10 was quantified by MB-AuNP-RT-I-PCR in EPTB cases, whereas magneto-ELISA demonstrated a narrow range (1.8-10 ng/ml). Furthermore, high sensitivity (88.2%) and specificity (100%) were attained by MB-AuNP-RT-I-PCR in EPTB (n = 51) and non-TB control (n = 49) subjects, respectively. Both MB-AuNP-I-PCR/magneto-ELISA exhibited significantly lower (p < 0.05-0.01) sensitivities than MB-AuNP-RT-I-PCR. The MB-AuNP-RT-I-PCR described herein shows good diagnostic accuracy, which may translate into a credible diagnostic kit.
肺外结核病（EPTB）的诊断较为困难，因此迫切需要一种快速可靠的诊断测试。研究了一种基于纳米的检测方法，即SYBR Green磁珠偶联金纳米颗粒实时免疫聚合酶链反应（MB-AuNP-RT-I-PCR），用于临床疑似EPTB患者中MPT-64+CFP-10蛋白的定量检测。在EPTB病例中，通过MB-AuNP-RT-I-PCR对宽范围（270 fg/ml - 9.9 ng/ml）的MPT-64+CFP-10进行了定量，而磁酶联免疫吸附测定（magneto-ELISA）显示的范围较窄（1.8 - 10 ng/ml）。此外，MB-AuNP-RT-I-PCR在EPTB患者（n = 51）和非结核对照组（n = 49）中分别达到了高灵敏度（88.2%）和特异性（100%）。与MB-AuNP-RT-I-PCR相比，MB-AuNP-I-PCR/磁酶联免疫吸附测定的灵敏度均显著降低（p < 0.05 - 0.01）。本文所述的MB-AuNP-RT-I-PCR显示出良好的诊断准确性，有望转化为一种可靠的诊断试剂盒。
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