Abstract

BackgroundCharacterization of the Mycobacterium leprae genome has made possible the development of Polymerase Chain Reaction (PCR) systems that can amplify different genomic regions. Increased reliability and technical efficiency of quantitative PCR (qPCR) makes it a promising tool for early diagnosis of leprosy. Index cases that are multibacillary spread the bacillus silently, even before they are clinically diagnosed. Early detection and treatment could prevent transmission in endemic areas.MethodsIn this study, the qPCR technique is used to detect DNA of M. leprae in samples of slit skin smears (SSS) of the ear lobe and blood of leprosy patients and their asymptomatic household contacts residing in Governador Valadares, MG, Brazil, a hyperendemic area for leprosy. A total of 164 subjects participated in the study: 43 index cases, 113 household contacts, and, as negative controls, 8 individuals who reported no contact with patients nor history of leprosy in the family. The qPCR was performed to amplify 16S rRNA fragments and was specifically designed for M. leprae.ResultsOf asymptomatic household contacts, 23.89% showed bacillary DNA by qPCR in samples of SSS and blood. Also, 48.84% of patients diagnosed with leprosy were positive for qPCR while the bacillary load was positive in only 30.23% of patients. It is important to note that most patients were already receiving treatment when the collection of biological material for qPCR was performed. The level of bacillary DNA from household contacts was similar to the DNA levels detected in the group of paucibacillary patients.ConclusionConsidering that household contacts comprise a recognizable group of individuals with a high risk of disease, as they live in close proximity to a source of infection, qPCR can be used to estimate the risk of progress towards leprosy among household contacts and as a routine screening method for a chemoprophylactic protocol.

Highlights

  • Characterization of the Mycobacterium leprae genome has made possible the development of Polymerase Chain Reaction (PCR) systems that can amplify different genomic regions

  • With respect to the MB group, it was found that 69.56% of patients were positive in both skin smears (SSS) or blood in quantitative PCR (qPCR) (Any-Sample)

  • It was observed that 30% of MB individuals that showed negative bacteriological index (BI) were qPCR positive

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Summary

Introduction

Characterization of the Mycobacterium leprae genome has made possible the development of Polymerase Chain Reaction (PCR) systems that can amplify different genomic regions. Increased reliability and technical efficiency of quantitative PCR (qPCR) makes it a promising tool for early diagnosis of leprosy. The early diagnosis and prompt initiation of treatment is essential to the rapid interruption of the disease transmission chain. In this sense, the development of a sensitive test for the diagnosis of leprosy has been one of the main objectives of research related to the disease [6]. The analysis of sensitivity and specificity of real-time quantitative PCR (qPCR) amplification of the sodA gene, 16S rRNA, RLEP, and 85BAg for the differential diagnosis of leprosy showed that the RLEP gene confers greater sensitivity ( a lower specificity) to the technique. We screened and followed household contacts (HHCs) along with patients, testing the 16S rRNA qPCR assay to evaluate the presence of M. leprae DNA in patients and their asymptomatic HHCs

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