Comparative evaluation of PCR amplification of RLEP, 16S rRNA, rpoT and Sod A gene targets for detection of M. leprae DNA from clinical and environmental samples

Turankar, Ravindra P.; Pandey, Shradha; Lavania, Mallika; Singh, Itu; Nigam, A. N.; Darlong, Joydeepa; Darlong, F; Sengupta, Utpal

doi:10.1016/j.ijmyco.2014.11.062

Cited by 51 publications

(33 citation statements)

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“…The high sensitivity of qPCR in relation to the BI makes this an extremely important technique in supporting the clinical diagnosis, as reported by other authors [ 12 , 13 , 20 ]. In our study, we confirmed the significant potential of qPCR for M. leprae DNA detection in leprosy cases with negative BI as well as in asymptomatic HHCs.…”

Section: Discussionmentioning

confidence: 71%

“…PCR is also a sensitive technique capable of detecting 25 fg (10 − 15 g) of M. leprae DNA [ 7 , 8 ]. The assays have been developed for regions such as the 36-kDa [ 9 ], 18-kDa [ 10 ] or 65-kDa [ 11 ] antigens as well as repeat sequences (RLEP) and the gene encoding the 16S rRNA of M. leprae [ 12 , 13 ]. 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 (although a lower specificity) to the technique.…”

Section: Introductionmentioning

confidence: 99%

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High frequency of M. leprae DNA detection in asymptomatic household contacts

et al. 2018

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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.Electronic supplementary materialThe online version of this article (10.1186/s12879-018-3056-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 71%

Section: Introductionmentioning

confidence: 99%

High frequency of M. leprae DNA detection in asymptomatic household contacts

et al. 2018

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“…Skin slit smears were taken from the patients from the border of their ear lobules according to Kamble et al [8] . Collected blood samples (five milliliters of venous blood) had been drawn into vacutainers tubes containing no anticoagulant.…”

Section: Inclusion Criteriamentioning

confidence: 99%

Polymerase Chain Reaction [PCR] versus Slit Skin Smear in Diagnosis of Leprosy

Kamel

Hashem

Khodair

et al. 2020

International Journal of Medical Arts

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Background: Leprosy is a chronic infectious disease where delay in diagnosis or treatment can lead to deformities and disabilities for the rest of the patient's life. Definite diagnosis of leprosy has long been based on clinical picture, histopathology and/or the detection of acid-fast bacilli (AFB) from tissue smears or tissue sections stained by Ziehl-Neelsen. Aim of the work: We evaluated in this study, the usefulness of PCR for detection of M.leprae in the serum, as less invasive technique, in comparison to skin slit smear for the diagnosis of leprosy. Patients and Methods: This study included 30 non-treated patients of leprosy (9 multibacillary, 21 paucibacillary) skin slit smears had been taken. In addition, serum samples had been collected for polymerase chain reaction [PCR] test. Results: Negative Slit Skin Smear for AFB had been obtained in 20 cases with a rate of positivity of 33%. Serum PCR was positive in 20 patients giving an overall rate of positivity of 67% which confirm the diagnosis in 11 out of 20 cases with slit skin smear [SSS] negative. Conclusion: PCR does show to be more sensitive than slit skin smear [SSS] indicating its future use for diagnostic purposes especially in early leprosy cases.

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“…Las herramientas genéticas como la RPC y la secuenciación genética, posibles después de la completa caracterización del genoma de M. leprae el 2011 20 , han revolucionado el conocimiento sobre la EH, permitiendo en algunos pacientes la confirmación en caso de duda diagnóstica, el conocimiento sobre sus mecanismos de transmisión, rol del reservorio humano y zoonótico, respuesta a la terapia y la identificación de mecanismos de resistencia al tratamiento [21][22][23][24][25][26][27] . Más aún, estas tecnologías han sido aplicadas tanto en muestras frescas como en muestras fijadas en parafina.…”

Section: Caso Clínicounclassified

Enfermedad de Hansen. Una condición emergente en Chile

San-Martín

Carrasco

Fíca

et al. 2018

Rev. chil. infectol.

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Hansen disease, an emergent condition in Chile Hansen's disease (HD) is caused by Mycobacterium leprae. It has a chronic course and preferentially affects the skin and the peripheral nerves. It´s an emergent infection in Chile due to migration waves. This case report affecting a migrant worker from Haiti that presented several compatible skin lesions, with hypoesthesia and unilateral madarosis that appeared before arrival. The diagnosis of a multibacillary form was established by clinical findings, presence of fast acid bacilli on a direct skin smear, and inflammatory cell surrounding nerve endings and granulomas on skin biopsy. Besides, specific rpoB and hsp65 gene segments from M. leprae were amplified from skin samples. Patient was treated with the WHO standard combined regimen for multibacillary forms during one year showing partial regression of skin lesions. Nasopharyngeal samples showed the presence of M. leprae rpoB copies detected by PCR decreasing until six months of therapy. Notifiable diseases databases showed a recent increment of cases, all related to migrant population. Hansen´s disease is a new condition in Chile and clinicians should be aware of this possibility. Molecular tools may facilitate diagnosis and follow up.

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Comparative evaluation of PCR amplification of RLEP, 16S rRNA, rpoT and Sod A gene targets for detection of M. leprae DNA from clinical and environmental samples

Cited by 51 publications

References 19 publications

High frequency of M. leprae DNA detection in asymptomatic household contacts

High frequency of M. leprae DNA detection in asymptomatic household contacts

Polymerase Chain Reaction [PCR] versus Slit Skin Smear in Diagnosis of Leprosy

Enfermedad de Hansen. Una condición emergente en Chile

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