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Reference Manager
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-
1.
Rao Pn, Suneetha S, 2018. Current situation of leprosy in India and its future implications. Indian Dermatol Online J 9: 83.
-
2.
Central Leprosy Division , 2016. National Leprosy Eradication Programme Annual Report 2015–2016. New Delhi, India: CLD.
-
3.
Central Leprosy Division , 2017. National Leprosy Eradication Programme Annual Report 2016–2017. New Delhi, India: CLD.
-
4.
Martinez AN, Britto CFPC, Nery JAC, Sampaio EP, Jardim MR, Sarno EN, Moraes MO, 2006. Evaluation of real-time and conventional PCR targeting complex 85 genes for detection of Mycobacterium leprae DNA in skin biopsy samples from patients diagnosed with leprosy. J Clin Microbiol 44: 3154–3159.
-
5.
Truman RW, Andrews PK, Robbins NY, Adams LB, Krahenbuhl JL, Gillis TP, 2008. Enumeration of Mycobacterium leprae using real-time PCR. PLoS Negl Trop Dis 2: e328.
-
6.
Liu Y-Y, Yu M-W, Ning Y, Wang H, 2018. A study on gender differences in newly detected leprosy cases in Sichuan, China, 2000–2015. Int J Dermatol 57: 1492–1499.
-
7.
Kumar A, Girdhar A, Girdhar BK, 2012. Six months fixed duration multidrug therapy in paucibacillary leprosy: risk of relapse and disability in Agra PB cohort study. BMJ Open 2: e001403.
-
8.
Bang PD, Suzuki K, Phuong LT, Chu TM, Ishii N, Khang TH, 2009. Evaluation of polymerase chain reaction-based detection of Mycobacterium leprae for the diagnosis of leprosy. J Dermatol 36: 269–276.
-
9.
Sarkar R, Pradhan S, 2016. Leprosy and women. Int J Womens Dermatol 2: 117–121.
-
10.
Siwakoti S, Rai K, Bhattarai NR, Agarwal S, Khanal B, 2016. Evaluation of polymerase chain reaction (PCR) with slit skin smear examination (SSS) to confirm clinical diagnosis of leprosy in eastern Nepal. PLoS Negl Trop Dis 10: e0005220.
-
11.
Demsiss W, Van Henten S, Takarinda KC, Kamau EM, Abdela SG, 2022. Slit-skin smear for the classification of leprosy; are we wasting time and resource? J Infect Dev Ctries 16: 3S–7S.
-
12.
Torres P, Camarena JJ, Gomez JR, Nogueira JM, Gimeno V, Navarro JC, Olmos A, 2003. Comparison of PCR mediated amplification of DNA and the classical methods for detection of Mycobacterium leprae in different types of clinical samples in leprosy patients and contacts. Leprosy 74: 18–30.
-
13.
Rudeeaneksin J, Srisungngam S, Sawanpanyalert P, Sittiwakin T, Likanonsakul S, Pasadorn S, Palittapongarnpim P, Brennan PJ, Phetsuksiri B, 2008. LightCyclerTM real-time PCR for rapid detection and quantitation of Mycobacterium leprae in skin specimens. FEMS Immunol Med Microbiol 54: 263–270.
-
14.
Yoon KH, Cho SN, Lee MK, Abalos RM, Cellona RV, Fajardo TT Jr. , Guido LS, Dela Cruz EC, Walsh GP, Kim JD, 1993. Evaluation of polymerase chain reaction amplification of Mycobacterium leprae-specific repetitive sequence in biopsy specimens from leprosy patients. J Clin Microbiol 31: 895–899.
-
15.
Azevedo MCS, Ramuno NM, Fachin LR, Tassa M, Rosa PS, Belone AF, Diório SM, Soares CT, Garlet GP, Trombone AP, 2017. qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms. Braz J Infect Dis 21: 71–78.
-
16.
Banerjee S, Biswas N, Kanti Das N, Sil A, Ghosh P, Hasanoor Raja AH, Dasgupta S, Kanti Datta P, Bhattacharya B, 2011. Diagnosing leprosy: revisiting the role of the slit-skin smear with critical analysis of the applicability of polymerase chain reaction in diagnosis: PCR versus slit-skin smear in diagnosis of leprosy. Int J Dermatol 50: 1522–1527.
-
17.
Goulart IMB, Cardoso AM, Santos MS, Gonçalves MA, Pereira JE, Goulart LR, 2007. Detection of Mycobacterium leprae DNA in skin lesions of leprosy patients by PCR may be affected by amplicon size. Arch Dermatol Res 299: 267–271.
-
18.
Reja AH, Biswas N, Biswas S, Dasgupta S, Chowdhury IH, Banerjee S, Chakraborty T, Dutta PK, Bhattacharya B, 2013. Fite–Faraco staining in combination with multiplex polymerase chain reaction: a new approach to leprosy diagnosis. Indian J Dermatol Venereol Leprol 79: 693–700.
-
19.
Kramme S, Bretzel G, Panning M, Kawuma J, Drosten C, 2004. Detection and quantification of Mycobacterium leprae in tissue samples by real-time PCR. Med Microbiol Immunol (Berl) 193: 189–193.
-
20.
Yan W, Xing Y, Yuan LC, Tan FY, De Yang R, Zhang Y, Li H-Y, 2014. Application of RLEP real-time PCR for detection of M. leprae DNA in paraffin-embedded skin biopsy specimens for diagnosis of paucibacillary leprosy. Am J Trop Med Hyg 90: 524–529.
-
21.
Lini N, Shankernarayan NP, Dharmalingam K, 2009. Quantitative real-time PCR analysis of Mycobacterium leprae DNA and mRNA in human biopsy material from leprosy and reactional cases. J Med Microbiol 58: 753–759.
| Past two years | Past Year | Past 30 Days | |
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Quantitative Real-Time Polymerase Chain Reaction for Detection of Mycobacterium leprae DNA in Tissue Specimens from Patients with Leprosy
Indukumari M. Sarath
Indukumari M. Sarath
Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education & Research, Puducherry, India
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Noyal Mariya Joseph
Noyal Mariya Joseph
Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education & Research, Puducherry, India
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Imola Jamir
Imola Jamir
Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education & Research, Puducherry, India
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ABSTRACT.
In leprosy, early diagnosis is crucial to prevent transmission and onset of disabilities of the disease. The purpose of this study was to determine usefulness of quantitative real-time polymerase chain reaction (PCR) in clinically diagnosed cases of leprosy. Thirty-two leprosy cases were included. The real-time PCR was performed using commercial kit targeting Mycobacterium leprae–specific insertion sequence element. The slit skin smear was positive in two (22.2%) borderline tuberculoid (BT) patients, five (83.3%) borderline lepromatous (BL) patients, and seven (50%) lepromatous leprosy (LL). The positivity of quantitative real-time PCR in BT, BL, LL, and pure neuritic leprosy were 77.8%, 83.3%, 100%, and 33.3%, respectively. Using histopathology as the gold standard, sensitivity of quantitative real-time PCR was 93.1%, and specificity was 100%. The DNA load was higher in LL (3,854.29/106 cells), followed by BL (140.37/106 cells), and BT (2.69/106 cells). Because of the high sensitivity and specificity of real-time PCR, our study strongly suggests the use of real-time PCR as a diagnostic tool for leprosy.
Author Notes
Financial support:
Authors’ addresses: Indukumari M. Sarath, Noyal Mariya Joseph, and Imola Jamir, Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India; E-mails: sarathmadhui@gmail.com, noyaljoseph@yahoo.com, and imolajamir3@gmail.com.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Rao Pn, Suneetha S, 2018. Current situation of leprosy in India and its future implications. Indian Dermatol Online J 9: 83.
-
2.
Central Leprosy Division , 2016. National Leprosy Eradication Programme Annual Report 2015–2016. New Delhi, India: CLD.
-
3.
Central Leprosy Division , 2017. National Leprosy Eradication Programme Annual Report 2016–2017. New Delhi, India: CLD.
-
4.
Martinez AN, Britto CFPC, Nery JAC, Sampaio EP, Jardim MR, Sarno EN, Moraes MO, 2006. Evaluation of real-time and conventional PCR targeting complex 85 genes for detection of Mycobacterium leprae DNA in skin biopsy samples from patients diagnosed with leprosy. J Clin Microbiol 44: 3154–3159.
-
5.
Truman RW, Andrews PK, Robbins NY, Adams LB, Krahenbuhl JL, Gillis TP, 2008. Enumeration of Mycobacterium leprae using real-time PCR. PLoS Negl Trop Dis 2: e328.
-
6.
Liu Y-Y, Yu M-W, Ning Y, Wang H, 2018. A study on gender differences in newly detected leprosy cases in Sichuan, China, 2000–2015. Int J Dermatol 57: 1492–1499.
-
7.
Kumar A, Girdhar A, Girdhar BK, 2012. Six months fixed duration multidrug therapy in paucibacillary leprosy: risk of relapse and disability in Agra PB cohort study. BMJ Open 2: e001403.
-
8.
Bang PD, Suzuki K, Phuong LT, Chu TM, Ishii N, Khang TH, 2009. Evaluation of polymerase chain reaction-based detection of Mycobacterium leprae for the diagnosis of leprosy. J Dermatol 36: 269–276.
-
9.
Sarkar R, Pradhan S, 2016. Leprosy and women. Int J Womens Dermatol 2: 117–121.
-
10.
Siwakoti S, Rai K, Bhattarai NR, Agarwal S, Khanal B, 2016. Evaluation of polymerase chain reaction (PCR) with slit skin smear examination (SSS) to confirm clinical diagnosis of leprosy in eastern Nepal. PLoS Negl Trop Dis 10: e0005220.
-
11.
Demsiss W, Van Henten S, Takarinda KC, Kamau EM, Abdela SG, 2022. Slit-skin smear for the classification of leprosy; are we wasting time and resource? J Infect Dev Ctries 16: 3S–7S.
-
12.
Torres P, Camarena JJ, Gomez JR, Nogueira JM, Gimeno V, Navarro JC, Olmos A, 2003. Comparison of PCR mediated amplification of DNA and the classical methods for detection of Mycobacterium leprae in different types of clinical samples in leprosy patients and contacts. Leprosy 74: 18–30.
-
13.
Rudeeaneksin J, Srisungngam S, Sawanpanyalert P, Sittiwakin T, Likanonsakul S, Pasadorn S, Palittapongarnpim P, Brennan PJ, Phetsuksiri B, 2008. LightCyclerTM real-time PCR for rapid detection and quantitation of Mycobacterium leprae in skin specimens. FEMS Immunol Med Microbiol 54: 263–270.
-
14.
Yoon KH, Cho SN, Lee MK, Abalos RM, Cellona RV, Fajardo TT Jr. , Guido LS, Dela Cruz EC, Walsh GP, Kim JD, 1993. Evaluation of polymerase chain reaction amplification of Mycobacterium leprae-specific repetitive sequence in biopsy specimens from leprosy patients. J Clin Microbiol 31: 895–899.
-
15.
Azevedo MCS, Ramuno NM, Fachin LR, Tassa M, Rosa PS, Belone AF, Diório SM, Soares CT, Garlet GP, Trombone AP, 2017. qPCR detection of Mycobacterium leprae in biopsies and slit skin smear of different leprosy clinical forms. Braz J Infect Dis 21: 71–78.
-
16.
Banerjee S, Biswas N, Kanti Das N, Sil A, Ghosh P, Hasanoor Raja AH, Dasgupta S, Kanti Datta P, Bhattacharya B, 2011. Diagnosing leprosy: revisiting the role of the slit-skin smear with critical analysis of the applicability of polymerase chain reaction in diagnosis: PCR versus slit-skin smear in diagnosis of leprosy. Int J Dermatol 50: 1522–1527.
-
17.
Goulart IMB, Cardoso AM, Santos MS, Gonçalves MA, Pereira JE, Goulart LR, 2007. Detection of Mycobacterium leprae DNA in skin lesions of leprosy patients by PCR may be affected by amplicon size. Arch Dermatol Res 299: 267–271.
-
18.
Reja AH, Biswas N, Biswas S, Dasgupta S, Chowdhury IH, Banerjee S, Chakraborty T, Dutta PK, Bhattacharya B, 2013. Fite–Faraco staining in combination with multiplex polymerase chain reaction: a new approach to leprosy diagnosis. Indian J Dermatol Venereol Leprol 79: 693–700.
-
19.
Kramme S, Bretzel G, Panning M, Kawuma J, Drosten C, 2004. Detection and quantification of Mycobacterium leprae in tissue samples by real-time PCR. Med Microbiol Immunol (Berl) 193: 189–193.
-
20.
Yan W, Xing Y, Yuan LC, Tan FY, De Yang R, Zhang Y, Li H-Y, 2014. Application of RLEP real-time PCR for detection of M. leprae DNA in paraffin-embedded skin biopsy specimens for diagnosis of paucibacillary leprosy. Am J Trop Med Hyg 90: 524–529.
-
21.
Lini N, Shankernarayan NP, Dharmalingam K, 2009. Quantitative real-time PCR analysis of Mycobacterium leprae DNA and mRNA in human biopsy material from leprosy and reactional cases. J Med Microbiol 58: 753–759.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1243 | 699 | 69 |
| Full Text Views | 262 | 215 | 0 |
| PDF Downloads | 59 | 30 | 0 |