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

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 Mycobacterium tuberculosis (MTB) DNA. The study involved testing whole blood samples for an MTB DNA target (known as IS6110). 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.

Keywords: Droplet Digital PCR; Mycobacterium tuberculosis; molecular diagnosis; tuberculosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cell-Free Nucleic Acids / blood*
  • DNA, Bacterial / blood*
  • Female
  • Humans
  • Male
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / isolation & purification*
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Tuberculosis, Pulmonary / blood
  • Tuberculosis, Pulmonary / diagnosis*

Substances

  • Cell-Free Nucleic Acids
  • DNA, Bacterial