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使用数字液滴PCR检测肺结核和肺外结核患者全血来源DNA样本中的DNA

Use of Digital Droplet PCR to Detect DNA in Whole Blood-Derived DNA Samples from Patients with Pulmonary and Extrapulmonary Tuberculosis.

作者信息

Yang Jiaru, Han Xinlin, Liu Aihua, Bai Xiyuan, Xu Cuiping, Bao Fukai, Feng Shi, Tao Lvyan, Ma Mingbiao, Peng Yun

机构信息

Yunnan Key Laboratory for Tropical Infectious DiseasesKunming, China.

Yunnan Collaborative Innovation Center for Public Health and Disease ControlKunming, China.

出版信息

Front Cell Infect Microbiol. 2017 Aug 11;7:369. doi: 10.3389/fcimb.2017.00369. eCollection 2017.

Abstract

Tuberculosis (TB) is a chronic infectious disease that has been threatening public health for many centuries. The clinical diagnostic procedure for TB is time-consuming and laborious. In the last 20 years, real-time fluorescence-based quantitative PCR (real-time PCR) has become a better alternative for TB diagnosis in clinics due to its sensitivity and specificity. Recently, digital droplet PCR (ddPCR) has been developed, and it might be an ideal alternative to conventional real-time PCR for microorganism detection. In this study, we aimed to assess the capacity of ddPCR and real-time PCR for detecting low levels of circulating (MTB) DNA. The study involved testing whole blood samples for an MTB DNA target (known as IS). Blood samples were obtained from 28 patients with pulmonary TB, 28 patients with extrapulmonary TB, and 28 healthy individuals. The results show that ddPCR could be used to measure low levels of MTB DNA, and it has the potential to be used to diagnose pulmonary and extrapulmonary TB based on clinical samples.

摘要

结核病(TB)是一种慢性传染病,几个世纪以来一直威胁着公众健康。结核病的临床诊断程序耗时且费力。在过去20年中,基于实时荧光的定量聚合酶链反应(实时PCR)因其灵敏度和特异性,已成为临床上结核病诊断的更好选择。最近,数字液滴PCR(ddPCR)已被开发出来,它可能是传统实时PCR用于微生物检测的理想替代方法。在本研究中,我们旨在评估ddPCR和实时PCR检测低水平循环结核分枝杆菌(MTB)DNA的能力。该研究涉及检测全血样本中的MTB DNA靶标(称为IS)。血液样本取自28例肺结核患者、28例肺外结核患者和28名健康个体。结果表明,ddPCR可用于测量低水平的MTB DNA,并且有潜力基于临床样本用于诊断肺结核和肺外结核。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71ee/5554497/339468340fcf/fcimb-07-00369-g0001.jpg

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