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-
1.
Foss NT, Motta ACF, 2012. Leprosy, a neglected disease that causes a wide variety of clinical conditions in tropical countries. Mem Inst Oswaldo Cruz, Rio de Janeiro 107 (Suppl 1): 28–33.
-
2.
Shepard CC, McRae DH, 1968. A method for counting acid-fast bacteria. Int J Lepr 36: 78–82.
-
3.
Buhrer-Sekula S, Smits HL, Gussenhoven GC, van Leeuwen J, Amador S, Fujiwara T, Klatser PR, Oskam L, 2003. Simple and fast lateral flow test for classification of leprosy patients and identification of contacts with high risk of developing leprosy. J Clin Microbiol 41: 1991–1995.
-
4.
Douglas JT, Steven LM, Fajardo T, Cellona RV, Madarang MG, Abalos RM, Steenbergen GJ, 1988. The effects of chemotherapy on antibody levels in lepromatous patients. Lepr Rev 59: 127–135.
-
5.
Santos VS, de Mendonça Neto PT, Falcão Raposo OF, Fakhouri R, Reis FP, Feitosa VL, 2013. Evaluation of agreement between clinical and histopathological data for classifying leprosy. Int J Infect Dis 17: e189–e192.
-
6.
Clark-Curtiss JE, Docherty MA, 1989. A species-specific repetitive sequence in Mycobacterium leprae DNA. J Infect Dis 159: 7–15.
-
7.
Katoch VM, 2001. Newer diagnostic techniques for tuberculosis. Indian J Med Res 120: 418–428.
-
8.
Hampson SJ, Portaels F, Thompson J, Green EP, Moss MT, Hermon-Taylor J, McFadden JJ, 1989. DNA probes demonstrate a single highly conserved strain of Mycobacterium avium infecting AIDS patients. Lancet 1: 65–68.
-
9.
Yoon D-H, Cho S-N, 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.
-
10.
Martinez AN, Ribeiro-Alves M, Sarno EN, Moraes MO, 2011. Evaluation of qPCR-based assays for leprosy diagnosis directly in clinical specimens. PLoS Negl Trop Dis 5: e1354.
-
11.
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.
-
12.
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.
-
13.
Hardman WJ, Benian GM, Howard T, McGowan JE Jr, Metchock B, Murtagh JJ, 1996. Rapid detection of mycobacteria in inflammatory necrotizing granulomas from formalin fixed, paraffin-embedded tissue by PCR in clinically high-risk patients with acid-fast stain and culture-negative tissue biopsies. Am J Clin Pathol 106: 384–389.
-
14.
Wen Y, Xing Y, Yuan LC, Liu J, Zhang Y, Li HY, 2013. Whole-blood nested-PCR amplification of M. leprae-specific DNA for early diagnosis of leprosy. Am J Trop Med Hyg 88: 918–922.
-
15.
Ridley DS, Jopling WH, 1966. Classification of leprosy according to immunity – a five group system. Int J Lepr 34: 255–273.
-
16.
Lockwood DN, Nicholls P, Smith WC, Das L, Barkataki P, van Brakel W, Suneetha S, 2012. Comparing the clinical and histological diagnosis of leprosy and leprosy reactions in the INFIR cohort of Indian patients with multibacillary leprosy. PLoS Negl Trop Dis 6: e1702.
-
17.
Cole ST, Supply P, Honoré N, 2001. Repetitive sequences in Mycobacterium leprae and their impact on genome plasticity. Lepr Rev 72: 449–461.
-
18.
Rodrigues LC, Lockwood DN, 2011. Leprosy now: epidemiology, progress, challenges, and research gaps. Lancet Infect Dis 11: 464–470.
-
19.
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.
-
20.
Scollard DM, Adams LB, Gillis TP, Kranenbuthl JL, Truman RW, Williams DL, 2006. The continuing challenges of leprosy. Clin Microbiol Rev 19: 338–381.
-
21.
Bhatia AS, Katoch K, Narayanan RB, Ramu G, Mukherjee A, Lavania RK, 1993. Clinical and histopathological correlation in the classification of leprosy. Int J Lepr 61: 433–438.
-
22.
Martinez AN, Britto CF, Nery JA, 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.
-
23.
Patrocínio LG, Goulart IM, Goulart LR, Patrocínio JA, Ferreira FR, Fleury RN, 2005. Detection of Mycobacterium leprae in nasal mucosa biopsies by the polymerase chain reaction. FEMS Immunol Med Microbiol 44: 311–316.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1268 | 974 | 21 |
| Full Text Views | 467 | 10 | 0 |
| PDF Downloads | 228 | 14 | 0 |
Application of RLEP Real-Time PCR for Detection of M. leprae DNA in Paraffin-Embedded Skin Biopsy Specimens for Diagnosis of Paucibacillary Leprosy
Wen Yan
Wen Yan
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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Yan Xing
Yan Xing
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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Lian Chao Yuan
Lian Chao Yuan
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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Rong De Yang
Rong De Yang
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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Fu Yue Tan
Fu Yue Tan
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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Ying Zhang
Ying Zhang
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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Huan-Ying Li
Huan-Ying Li
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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The TaqMan real-time polymerase chain reaction (PCR) assay was evaluated systematically with respect to the standard curve, linear range, and used for detecting Mycobacterium leprae DNA in paraffin-embedded skin biopsy specimens from 60 confirmed leprosy patients and three healthy individuals and 29 other dermatoses and bacterial DNA from 21 different species. The test was further evaluated with 51 paucibacillary (PB) patients. The results showed that the test had good sensitivity (8 fg) and good specificity with no cross-reactivity with 21 other bacterial species and the control specimens, except one with Xanthomatosis. The real-time PCR detection rate for the 51 PB specimens was 74.5% (38 of 51). We conclude that the real-time PCR test is a useful adjunct test for diagnosing early stage or PB leprosy cases.
Author Notes
Authors' addresses: Wen Yan, Yan Xing, Lian Chao Yuan, and Huan-Ying Li, Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing, China, E-mails: weny8@163.com, hellen19801@163.com, yuanlc@126.com, and lhybj@163.com. Rong De Yang and Fu Yue Tan, Yunnan Wenshan Skin Disease Dispensary, China, E-mails: yangrd@qq.com and tfyyyy@qq.com Ying Zhang, Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, E-mail: yzhang@jhsph.edu.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Foss NT, Motta ACF, 2012. Leprosy, a neglected disease that causes a wide variety of clinical conditions in tropical countries. Mem Inst Oswaldo Cruz, Rio de Janeiro 107 (Suppl 1): 28–33.
-
2.
Shepard CC, McRae DH, 1968. A method for counting acid-fast bacteria. Int J Lepr 36: 78–82.
-
3.
Buhrer-Sekula S, Smits HL, Gussenhoven GC, van Leeuwen J, Amador S, Fujiwara T, Klatser PR, Oskam L, 2003. Simple and fast lateral flow test for classification of leprosy patients and identification of contacts with high risk of developing leprosy. J Clin Microbiol 41: 1991–1995.
-
4.
Douglas JT, Steven LM, Fajardo T, Cellona RV, Madarang MG, Abalos RM, Steenbergen GJ, 1988. The effects of chemotherapy on antibody levels in lepromatous patients. Lepr Rev 59: 127–135.
-
5.
Santos VS, de Mendonça Neto PT, Falcão Raposo OF, Fakhouri R, Reis FP, Feitosa VL, 2013. Evaluation of agreement between clinical and histopathological data for classifying leprosy. Int J Infect Dis 17: e189–e192.
-
6.
Clark-Curtiss JE, Docherty MA, 1989. A species-specific repetitive sequence in Mycobacterium leprae DNA. J Infect Dis 159: 7–15.
-
7.
Katoch VM, 2001. Newer diagnostic techniques for tuberculosis. Indian J Med Res 120: 418–428.
-
8.
Hampson SJ, Portaels F, Thompson J, Green EP, Moss MT, Hermon-Taylor J, McFadden JJ, 1989. DNA probes demonstrate a single highly conserved strain of Mycobacterium avium infecting AIDS patients. Lancet 1: 65–68.
-
9.
Yoon D-H, Cho S-N, 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.
-
10.
Martinez AN, Ribeiro-Alves M, Sarno EN, Moraes MO, 2011. Evaluation of qPCR-based assays for leprosy diagnosis directly in clinical specimens. PLoS Negl Trop Dis 5: e1354.
-
11.
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.
-
12.
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.
-
13.
Hardman WJ, Benian GM, Howard T, McGowan JE Jr, Metchock B, Murtagh JJ, 1996. Rapid detection of mycobacteria in inflammatory necrotizing granulomas from formalin fixed, paraffin-embedded tissue by PCR in clinically high-risk patients with acid-fast stain and culture-negative tissue biopsies. Am J Clin Pathol 106: 384–389.
-
14.
Wen Y, Xing Y, Yuan LC, Liu J, Zhang Y, Li HY, 2013. Whole-blood nested-PCR amplification of M. leprae-specific DNA for early diagnosis of leprosy. Am J Trop Med Hyg 88: 918–922.
-
15.
Ridley DS, Jopling WH, 1966. Classification of leprosy according to immunity – a five group system. Int J Lepr 34: 255–273.
-
16.
Lockwood DN, Nicholls P, Smith WC, Das L, Barkataki P, van Brakel W, Suneetha S, 2012. Comparing the clinical and histological diagnosis of leprosy and leprosy reactions in the INFIR cohort of Indian patients with multibacillary leprosy. PLoS Negl Trop Dis 6: e1702.
-
17.
Cole ST, Supply P, Honoré N, 2001. Repetitive sequences in Mycobacterium leprae and their impact on genome plasticity. Lepr Rev 72: 449–461.
-
18.
Rodrigues LC, Lockwood DN, 2011. Leprosy now: epidemiology, progress, challenges, and research gaps. Lancet Infect Dis 11: 464–470.
-
19.
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.
-
20.
Scollard DM, Adams LB, Gillis TP, Kranenbuthl JL, Truman RW, Williams DL, 2006. The continuing challenges of leprosy. Clin Microbiol Rev 19: 338–381.
-
21.
Bhatia AS, Katoch K, Narayanan RB, Ramu G, Mukherjee A, Lavania RK, 1993. Clinical and histopathological correlation in the classification of leprosy. Int J Lepr 61: 433–438.
-
22.
Martinez AN, Britto CF, Nery JA, 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.
-
23.
Patrocínio LG, Goulart IM, Goulart LR, Patrocínio JA, Ferreira FR, Fleury RN, 2005. Detection of Mycobacterium leprae in nasal mucosa biopsies by the polymerase chain reaction. FEMS Immunol Med Microbiol 44: 311–316.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1268 | 974 | 21 |
| Full Text Views | 467 | 10 | 0 |
| PDF Downloads | 228 | 14 | 0 |