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-
1.
Duthie MS, Goto W, Ireton GC, Reece ST, Cardoso LP, Martelli CM, Stefani MM, Nakatani M, de Jesus RC, Netto EM, Balagon MV, Tan E, Gelber RH, Maeda Y, Makino M, Hoft D, Reed SG, 2007. Use of protein antigens for early serological diagnosis of leprosy. Clin Vaccine Immunol 14: 1400–1408.
-
2.
Douglas JT, Worth RM, 1984. Field evaluation of an ELISA to detect antibody in leprosy patients and their contacts. Int J Lepr Other Mycobact Dis 52: 26–33.
-
3.
De Wit MYL, Faber WR, Krieg SR, Douglas JT, Lucas SB, Montreewasuwat N, Pattyn SR, Hussain R, Ponnighaus JM, Hartskeerl RA, 1991. Application of a polymerase chain reaction for the detection of Mycobacterium leprae in skin tissues. J Clin Microbiol 29: 906–910.
-
4.
Woods SA, Cole ST, 1989. A rapid method for the detection of potentially viable Mycobacterium leprae in human biopsies: a novel application of PCR. FEMS Microbiol Lett 65: 305–310.
-
5.
Plikaytis BB, Gelber RH, Shinnick TM, 1990. Rapid and sensitive detection of Mycobacterium leprae using a nested-primer gene amplification assay. J Clin Microbiol 28: 1913–1917.
-
6.
Wang HB, Weng XM, Wen Y, Li HY, 2005. Improved detection of Mycobacterium leprae rifampin resistance with nested PCR. Chin J Lab Med 28: 1015–1017.
-
7.
Ridley DS, Jopling WH, 1966. Classification of leprosy according to immunity—a five group system. Int J Lepr 34: 255–273.
-
8.
Wen Y, Vissa VD, Li W, Li HY, 2010. Antigenicity of Mycobacterium leprae recombinant proteins. J Trop Med 10: 1153–1157.
-
9.
Sreevatsa RG, Sengupta U, Ramu G, Desikan KV, 1978. Evaluation of bacteraemia in leprosy patients. Lepr India 50: 381–387.
-
10.
Santos AR, De Miranda AB, Sarno EN, Suffys PN, Degrave WM, 1993. Use of PCR-mediated amplification of Mycobacterium leprae DNA in different types of clinical samples for the diagnosis of leprosy. J Med Microbiol 39: 298–304.
-
11.
Kaur I, Kaur S, Sharma VK, Agnihotri N, Vaishnavi C, Ganguly NK, 1993. Bacillaemia and Mycobacterium leprae cell wall antigen in paucibacillary leprosy. Indian J Lepr 65: 283–288.
-
12.
Agusni I, Prakoeswa CR, Adriaty D, 2008. Detection of M. leprae in the blood of leprosy patients and healthy inhabitants live in endemic leprosy area. Proceedings of the 17th International Leprosy Congress, Hyderabad, India, 30th January–4th February 2008.
-
13.
Hartskeerl RA, De Wit MYL, Klatser PR, 1989. Polymerase chain reaction for the detection of Mycobacterium leprae. J Gen Microbiol 135: 2357–2364.
-
14.
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.
-
15.
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.
-
16.
Oskam L, Slim E, Buhrer-Se'kula S, 2003. Serology: recent developments, strengths, limitations and prospects: a state of the art overview. Lepr Rev 74: 196–205.
-
17.
Klatser PR, Cho SN, Brennan PJ, 1996. The contribution of serological tests to leprosy control. Int J Lepr Other Mycobact Dis 64 (Suppl 4): S63–S66.
-
18.
Menzel S, Harboe M, Bergsvik H, Brennan PJ, 1987. Antibodies to a synthetic analog of phenolic glycolipid-I of Mycobacterium leprae in healthy household contacts of patients with leprosy. Int J Lepr Other Mycobact Dis 55: 617–625.
-
19.
Saad MH, Medeiros MA, Gallo ME, Gontijo PP, Fonseca LS, 1990. IgM immunoglobulins reacting with the phenolic glycolipid-1 antigen from Mycobacterium leprae in sera of leprosy patients and their contacts. Mem Inst Oswaldo Cruz 85: 191–194.
-
20.
Banerjee S, Sarkar K, Gupta S, Mahapatra PS, Gupta S, Guha S, Bandhopadhayay D, Ghosal C, Paine SK, Dutta RN, Biswas N, Bhattacharya B, 2010. Multiplex PCR technique could be an alternative approach for early detection of leprosy among close contacts—a pilot study from India. BMC Infect Dis 10: 252.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1180 | 1060 | 626 |
| Full Text Views | 423 | 11 | 0 |
| PDF Downloads | 138 | 5 | 0 |
Whole-Blood Nested-PCR Amplification of M. leprae-Specific DNA for Early Diagnosis of Leprosy
Yan Wen
Yan Wen
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, 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; 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; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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Jian Liu
Jian Liu
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, 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; 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; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland
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We evaluated the sensitivity and specificity of a nested-polymerase chain reaction (PCR) method for detection of Mycobacterium leprae DNA from whole blood. Whole-blood specimens were subjected to nested-PCR amplification of M. leprae repeat DNA sequences in 49 multibacillary (MB) and 30 paucibacillary (PB) leprosy patients, 96 household contacts (HHCs), 18 tuberculosis (TB) patients, and 35 normal healthy individuals. M. leprae DNA was detected in 95.92% (47/49) of MB, 70% (21/30) of PB, and 6.25% (6/96) of HHC, but it was not detected in 18 TB or 35 normal controls. The sensitivities of the anti-bovine serum albumin (ND-O-BSA) immunoglobulin M (IgM) and antifusion protein of ML0405-ML2331 IgG for MB were 97.96% and 89.8%, and these values for PB were 70% and 53.33%. However, the ND-O-BSA enzyme-linked immunosorbent assay (ELISA) had lower specificity, with relatively high false-positive results for TB patients (16.67%) and normal healthy controls (10%). Based on these promising findings, we propose the use of nested PCR of whole-blood samples along with ELISA test for early detection of leprosy cases.
Author Notes
Authors' addresses: Yan Wen, Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China, E-mail: weny8@163.com. Yan Xing, Lian-Chao Yuan, Jian Liu, and Huan-Ying Li, Department of Leprosy Research, Beijing Tropical Medicine Research Institute, Beijing, China, E-mails: xingyan@163.com, yuanlianchao@126.com, liujian@sina.com, and lihybj@163.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.
Duthie MS, Goto W, Ireton GC, Reece ST, Cardoso LP, Martelli CM, Stefani MM, Nakatani M, de Jesus RC, Netto EM, Balagon MV, Tan E, Gelber RH, Maeda Y, Makino M, Hoft D, Reed SG, 2007. Use of protein antigens for early serological diagnosis of leprosy. Clin Vaccine Immunol 14: 1400–1408.
-
2.
Douglas JT, Worth RM, 1984. Field evaluation of an ELISA to detect antibody in leprosy patients and their contacts. Int J Lepr Other Mycobact Dis 52: 26–33.
-
3.
De Wit MYL, Faber WR, Krieg SR, Douglas JT, Lucas SB, Montreewasuwat N, Pattyn SR, Hussain R, Ponnighaus JM, Hartskeerl RA, 1991. Application of a polymerase chain reaction for the detection of Mycobacterium leprae in skin tissues. J Clin Microbiol 29: 906–910.
-
4.
Woods SA, Cole ST, 1989. A rapid method for the detection of potentially viable Mycobacterium leprae in human biopsies: a novel application of PCR. FEMS Microbiol Lett 65: 305–310.
-
5.
Plikaytis BB, Gelber RH, Shinnick TM, 1990. Rapid and sensitive detection of Mycobacterium leprae using a nested-primer gene amplification assay. J Clin Microbiol 28: 1913–1917.
-
6.
Wang HB, Weng XM, Wen Y, Li HY, 2005. Improved detection of Mycobacterium leprae rifampin resistance with nested PCR. Chin J Lab Med 28: 1015–1017.
-
7.
Ridley DS, Jopling WH, 1966. Classification of leprosy according to immunity—a five group system. Int J Lepr 34: 255–273.
-
8.
Wen Y, Vissa VD, Li W, Li HY, 2010. Antigenicity of Mycobacterium leprae recombinant proteins. J Trop Med 10: 1153–1157.
-
9.
Sreevatsa RG, Sengupta U, Ramu G, Desikan KV, 1978. Evaluation of bacteraemia in leprosy patients. Lepr India 50: 381–387.
-
10.
Santos AR, De Miranda AB, Sarno EN, Suffys PN, Degrave WM, 1993. Use of PCR-mediated amplification of Mycobacterium leprae DNA in different types of clinical samples for the diagnosis of leprosy. J Med Microbiol 39: 298–304.
-
11.
Kaur I, Kaur S, Sharma VK, Agnihotri N, Vaishnavi C, Ganguly NK, 1993. Bacillaemia and Mycobacterium leprae cell wall antigen in paucibacillary leprosy. Indian J Lepr 65: 283–288.
-
12.
Agusni I, Prakoeswa CR, Adriaty D, 2008. Detection of M. leprae in the blood of leprosy patients and healthy inhabitants live in endemic leprosy area. Proceedings of the 17th International Leprosy Congress, Hyderabad, India, 30th January–4th February 2008.
-
13.
Hartskeerl RA, De Wit MYL, Klatser PR, 1989. Polymerase chain reaction for the detection of Mycobacterium leprae. J Gen Microbiol 135: 2357–2364.
-
14.
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.
-
15.
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.
-
16.
Oskam L, Slim E, Buhrer-Se'kula S, 2003. Serology: recent developments, strengths, limitations and prospects: a state of the art overview. Lepr Rev 74: 196–205.
-
17.
Klatser PR, Cho SN, Brennan PJ, 1996. The contribution of serological tests to leprosy control. Int J Lepr Other Mycobact Dis 64 (Suppl 4): S63–S66.
-
18.
Menzel S, Harboe M, Bergsvik H, Brennan PJ, 1987. Antibodies to a synthetic analog of phenolic glycolipid-I of Mycobacterium leprae in healthy household contacts of patients with leprosy. Int J Lepr Other Mycobact Dis 55: 617–625.
-
19.
Saad MH, Medeiros MA, Gallo ME, Gontijo PP, Fonseca LS, 1990. IgM immunoglobulins reacting with the phenolic glycolipid-1 antigen from Mycobacterium leprae in sera of leprosy patients and their contacts. Mem Inst Oswaldo Cruz 85: 191–194.
-
20.
Banerjee S, Sarkar K, Gupta S, Mahapatra PS, Gupta S, Guha S, Bandhopadhayay D, Ghosal C, Paine SK, Dutta RN, Biswas N, Bhattacharya B, 2010. Multiplex PCR technique could be an alternative approach for early detection of leprosy among close contacts—a pilot study from India. BMC Infect Dis 10: 252.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1180 | 1060 | 626 |
| Full Text Views | 423 | 11 | 0 |
| PDF Downloads | 138 | 5 | 0 |