Dahiya Bhawna, Prasad Tulika, Rais Anam, Sheoran Abhishek, Kamra Ekta, Mor Preeti, Soni Aishwarya, Sharma Suman, Mehta Promod K
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.
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。
