Using of Digital Droplet PCR in the detection of Mycobacterium tuberculosis DNA with FFPE and Cytological samples

Abstract

Backgrounds: Accurate diagnosis for TB is essential for TB control. Droplet digital PCR (ddPCR) is a technology that has high sensitivity. However, the use of ddPCR for the detection of TB pathogen in pathological samples is not been fully studied.Methods: A total of 88 samples from the patients who were suspected of tuberculosis were involved in this study, including 65 formalin-fixed and paraffin-embedded (FFPE) specimens and 22 cytological samples. Digital Droplet PCR was used to compare the sensitivity with Real-time PCR. The real-time PCR ct value and ddPCR TB abundant ratio were analyzed by SPSS software.Results: Among the 62 samples that were “possible TB” detected by real-time PCR, 34 samples were ddPCR-positive, 18 samples were ddPCR-negative, and 10 samples were in “gray area” by ddPCR. 26 patients that were ddPCR-positive received anti-tuberculosis therapy and 14 cases of them benefit from the treatment.Conclusions: ddPCR is more sensitive in the detection of TB than Real-time PCR. ddPCR methods can be used as an additional means for the diagnosis of TB from pathological samples.