Nasal PCR assay for the detection of Mycobacterium leprae pra gene to study subclinical infection in a community

Arunagiri, Kavita; Gopalakrishnan, Subramaniam; Sugashini, Padmavathy Krishnan; Sekar, B; Ali, Mumtaz

doi:10.1016/j.micpath.2017.01.046

Cited by 11 publications

(10 citation statements)

References 6 publications

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“…In endemic regions, detection of M. leprae DNA from nasal secretions does not differentiate contacts from cases [16,19,35]; therefore, quantification of M. leprae DNA can be used in epidemiological studies to identify healthy carriers among household contacts or within populations of an endemic area [7,36].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of 16S rRNA qPCR for detection ofMycobacterium lepraeDNA in nasal secretion and skin biopsy samples from multibacillary and paucibacillary leprosy cases

Marques

Frota

Quetz

et al. 2017

Pathogens and Global Health

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Mycobacterium leprae bacilli are mainly transmitted by the dissemination of nasal aerosols from multibacillary (MB) patients to susceptible individuals through inhalation. The upper respiratory tract represents the main entry and exit routes of M. leprae. Therefore, this study aimed to evaluate the sensitivity and specificity of real-time quantitative polymerase chain reaction (qPCR) in detecting M. leprae in nasal secretion (NS) and skin biopsy (SB) samples from MB and paucibacillary (PB) cases. Fifty-four NS samples were obtained from leprosy patients at the Dona Libânia National Reference Centre for Sanitary Dermatology in Ceará, Brazil. Among them, 19 MB cases provided both NS and SB samples. Bacilloscopy index assays were conducted and qPCR amplification was performed using specific primers for M. leprae 16S rRNA gene, generating a 124-bp fragment. Primer specificity was verified by determining the amplicon melting temperature (T = 79.5 °C) and detection limit of qPCR was 20 fg of M. leprae DNA. Results were positive for 89.7 and 73.3% of NS samples from MB and PB cases, respectively. SB samples from MB patients were 100% positive. The number of bacilli detected in NS samples were 1.39 × 10-8.02 × 10, and in SB samples from MB patients were 1.87 × 10-1.50 × 10. Therefore, qPCR assays using SYBR Green targeting M. leprae 16S rRNA region can be employed in detecting M. leprae in nasal swabs from leprosy patients, validating this method for epidemiological studies aiming to identify healthy carriers among household contacts or within populations of an endemic area.

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Section: Discussionmentioning

confidence: 99%

Evaluation of 16S rRNA qPCR for detection ofMycobacterium lepraeDNA in nasal secretion and skin biopsy samples from multibacillary and paucibacillary leprosy cases

Marques

Frota

Quetz

et al. 2017

Pathogens and Global Health

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“…15 . So PCR of nasal smear in case of subclinical infection in a community could be a helpful diagnostic tool 16 . The PB type carries so few M. leprae organisms that these cases could not be detected by microscopic examination.…”

Section: Figures In Parenthesis Indicate Percentagementioning

confidence: 99%

Diagnosis of leprosy by PCR targeting gene encoding 36 kDa antigen of Mycobacterium leprae

Shampa

Roy

Begam

et al. 2020

Bangladesh J Med Microbiol

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Background & Objective: This cross sectional study was carried out to assess the diagnostic value of PCR in different forms of leprosy. For the detection of Mycobacterium leprae, DNA amplification by polymerase chain reaction of a 531-bp fragment of the Mycobacterium leprae specific gene encoding the 36 kDA antigen. Methodology: It was done on different clinical specimens (slit smear of skin, ear lobule smear and nasal smear) from 50 leprosy patients attending the Leprosy Hospital, Mohakhali, Dhaka. Patients were divided into two groups; paucibacillary (70%) group and multibacillary (30%) group. PCR showed 100% positivity in skin and ear lobule and 73.4% positivity in nasal smear of multibacillary group. PCR was positive in 40%, 25.7% and 11.4% in skin lesion, ear lobule and nasal swab in paucibacillry group respectively. Result: Compared with other diagnostic procedures, PCR showed clear advantages over both modified Z-N stain and auramine-phenol stain especially in paucibacillary patients. Bangladesh J Med Microbiol 2020; 14 (1): 11-14

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“…In the same way, the methodology can be useful, for those patients with the pure neural form (PNL), who usually do not have cutaneous lesions and because of this they have deficient treatment scheme [65,66]. Further, the PCR technique may be useful in early identification, since a considerable number of studies have addressed the positivity of M. leprae DNA in contacts of leprosy patients [48,[67][68][69].…”

Section: Complementary Molecular Testsmentioning

confidence: 99%

Molecular and Biotechnological Approaches in the Diagnosis of Leprosy

Lima¹,

Moraes²,

Pinho³

et al. 2019

Hansen's Disease - The Forgotten and Neglected Disease

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Leprosy is a worldwide health problem, which needs the development of new and innovative strategies to be controlled. Early diagnosis of leprosy is an important contribution to reducing the incidence of the disease; thus, the development of biotechnology platforms, which include the mapping of antigens with potential to be used in immunodiagnostic and molecular methods for the detection of Mycobacterium leprae, is an important tool to confirm the clinical diagnostic. Molecular biology and biotechnological methods have been used to assist in the diagnosis of this disease, each one with its advantages and drawbacks. Enzyme-linked immunosorbent assay (ELISA) is the used method for leprosy diagnosis, and it allows the detection of infection-related antigens. Alternatively, due to their versatility to perform the same functions as the protein and non-protein natural antigens, mimetic peptides are considered an important tool. On the other hand, lateral flow assay (LFA) and optical and electrochemical biosensors are rapid and portable methods, capable of performing diagnosis in the field without sample preparation.This chapter presents such techniques, their uses in the diagnosis and detection of M. leprae, as well as the potential for the development of new techniques and strategies that can help to control and understand mycobacteriosis.

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Nasal PCR assay for the detection of Mycobacterium leprae pra gene to study subclinical infection in a community

Cited by 11 publications

References 6 publications

Evaluation of 16S rRNA qPCR for detection ofMycobacterium lepraeDNA in nasal secretion and skin biopsy samples from multibacillary and paucibacillary leprosy cases

Evaluation of 16S rRNA qPCR for detection ofMycobacterium lepraeDNA in nasal secretion and skin biopsy samples from multibacillary and paucibacillary leprosy cases

Diagnosis of leprosy by PCR targeting gene encoding 36 kDa antigen of Mycobacterium leprae

Molecular and Biotechnological Approaches in the Diagnosis of Leprosy

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