Detection of Mycobacterium ulcerans by real-time PCR with improved primers. Issue 1 (December 2016)

• Record Type: Journal Article
• Authors: Shinoda, Noriko; Nakamura, Hajime; Watanabe, Mineo
• Is Part Of: Tropical medicine and health. Volume 44:Issue 1(2016)
• Page Start: 1
• Page End: 6
• Publication Date: 2016-12
• Subjects: Buruli ulcer -- Mycobacterium ulcerans -- Diagnosis -- Real-time PCR; 616.9883
• DOI: 10.1186/s41182-016-0028-3
• Languages: English
• ISSNs: 1349-4147

Abstract

Abstract Background Buruli ulcer is a severe skin disease caused byMycobacterium ulcerans . Real-time PCR targeting the IS2404 sequence has been used as a reliable and rapid method for the diagnosis of Buruli ulcer and detection ofM. ulcerans in the environment. The genome ofM. ulcerans contains hundreds of IS2404 copies, which have variability in certain sequences. Therefore, the design of new primers specific to conserved IS2404 regions may potentially improve the sensitivity ofM. ulcerans detection and, consequently, the diagnosis of Buruli ulcer, thus ensuring timely treatment of the disease. Results In silico analysis indicates that DNA sequences of the IS2404 elements are highly variable within a single strain. As the binding sites of conventional IS2404 -specific primers used forM. ulcerans detection contain polymorphic sequences, we designed new primers, which enabled the detection ofM. ulcerans by real-time PCR with higher sensitivity and similar specificity with respect to that of conventional primers. However, the increase in sensitivity with the new primers depended on theM. ulcerans strain. Conclusions The results suggest that real-time PCR based on the new primers could improve Buruli ulcer diagnosis andM. ulcerans detection in environmental samples.