Background: Nerve damage in leprosy reaction occurs very quickly, that can lead to paralysis for many peripheral nerves within 24 hours. Objective: To survey nerve function impairment in leprosy reactions. Subjects and method: 285 new leprosy patients treated from 1996 to 2000 at 22 districts in Ho Chi Minh city had leprosy reaction and estimated nerve functions. This was retrospective cross-section descriptive study. Results: This study included 70.2% men, age 15-45 years (71.9%) and most patients were manual laborers. There were 83.5% multibacillary (MB) and 16.5% paucibacillary (PB). 86.3% Reversal Reaction (RR), Erythemal Nodosum Leprosy (ENL) 13% and both 0.7%. 53% of leprosy reaction happened during Multi-Drug Treatment (MDT), at diagnosis 37.5%, before MDT 7.7% and after MDT 1.8%. Nerve function impairment was 38.2% in leprosy reaction; men had impaired nerve function 1.7 times higher than women (p<0.05). Nerve function impairment in RR and ENL as well as MB and PB were the same (p>0.05). MB (42%) has impaired nerve function higher than PB (26.1%) in RR (p<0.05, OR = 2.05). Conclusions: Leprosy reaction happened to men more frequent than women, essentially in MB and during MDT. One third of cases had nerve function impairment in leprosy reaction and also had a link between sex and nerve function impairment. MB was 2 times the impaired nerve function higher than PB in RR.
Nerve function impairment ; leprosy reaction

There are several methods for diagnosis of leprosy, including AFB stain, the measurement of PGL-1 (phenolic glycolipid - 1) antigen titer, and DNA-PCR. In this study, we have used the DNA-PCR amplifying the RLEP repetitive sequence. Our result showed that the RLEP primer offered the more sensitive detection and identification of M. leprae DNA in clinical specimens, compared with the other primer, for example, 18-kDa antigen gene. To screen the resistant M. leprae strain of MDT (Multi-Drug Therapy), we have used the TD (Touch-Down) PCR. We arranged and amplified sequences of the genes, folP, rpoB, gyr, 23S rRNA, in M. leprae involved in MDT-resistance, and could obtain the PCR product each gene, simultaneously. This method, based on annealing temperature, was useful to the detection for diagnosis and the screen of MDT-resistant strain of M. leprae, rapidly. Thus, we suggest that the RLEP primer and TD-PCR method are effective in assessing the diagnosis of leprosy and the identification of drug-resistant M. leprae.
Diagnosis* ; DNA ; Leprosy* ; Polymerase Chain Reaction* ; Repetitive Sequences, Nucleic Acid

BACKGROUND: The causative agents of leprosy are the well-known Mycobacterium leprae and the newly discovered Mycobacterium lepromatosis. This agent was found in 2008, and it was found to be the cause of diffuse lepromatous leprosy in two Mexican patients. OBJECTIVE: The objective of this work was to determine if M. leprae and M. lepromatosis were present in formalin-fixed and paraffin-embedded skin samples from cases from different regions in Mexico. METHODS: A total of 41 skin samples were obtained from 11 states of Mexico. All patients' samples were diagnosed by clinical and histopathological analyses. Total DNA was isolated using a Qiagen-DNeasy blood and tissue kit and molecular identification was achieved by two semi-nested polymerase chain reactions. RESULTS: The 41 patient included 33 samples from men and 8 samples from women; 29 samples were polymerase chain reaction (PCR)-positive to Mycobacterium and 12 samples were PCR-negative. From those 29 samples, 13 were PCR-positive to M. leprae, 8 to M. lepromatosis and 8 were positive to both species. The histopathological diagnosis included; Nodular lepromatous leprosy (NLL); Diffuse lepromatous leprosy (DLL); and Borderline leprosy (BL). The 29 PCR-positive samples were classified as follow: 14 NLL, 4 DLL, and 11 BL. In the 12 samples negative to Mycobacterium, 7 showed the NLL, 2 DLL and 3 BL. CONCLUSION: These findings add evidence to the M. leprae and M. lepromatous distribution, clinical forms and participation of dual infections in Mexico.
Diagnosis ; DNA ; Female ; Hospital Distribution Systems ; Humans ; Leprosy ; Leprosy, Borderline ; Leprosy, Lepromatous ; Male ; Mexico* ; Mycobacterium leprae* ; Mycobacterium* ; Polymerase Chain Reaction ; Skin*

BACKGROUND: Diagnosis of paucibacillary leprosy is difficult owing to lack of sensitive diagnostic tools. Recently, several investigators have studied the use of polymerase chain reaction(PCR) to detect Mycobacterium leprae. Using nested-PCR the sensitivity and specificity of DNA amplification is considerably improved. OBJECTIVE: The purpose of investigation is to assess the efficacy to nested-PCR which is applied to formalin-fixed, paraffin-embedded biopsies material of patients with leprosy. METHODS: Biopsy samples were taken from patients with lepromatous(11 patients) and tuberculoid (10 patients) leprosy, fixed in formalin, and embedded in paraffin. The DNA from samples was extracted and amplified through 25 cycles by using the outside pairs of primer(L1 and L2). The second amplification was allowed to proceed through 15 cycles using inside pairs of primer(L3 and L4). RESULTS: All twenty one samples showed 347-base-pair products. To confirm that the 347-bp product did correspond to the expected portion of the M. leprae groEL gene, the amplified product was digested with Pst I. Pst I digestion yielded 254-and 93-bp fragments, as predicted from the sequence of the M. leprae gene. The sensitivity was that a single organism was identified by nested-PCR. CONCLUSION: The nested-PCR is sensitive, specific, and simple diagnostic tool for leprosy.
Biopsy ; Diagnosis ; Digestion ; DNA* ; Formaldehyde ; Humans ; Leprosy ; Leprosy, Paucibacillary ; Mycobacterium leprae* ; Mycobacterium* ; Paraffin* ; Polymerase Chain Reaction* ; Research Personnel ; Sensitivity and Specificity

Leprosy is an important health problem worldwide yet. It is caused by a chronic granulomatous infection of the skin and peripheral nerves with Mycobacterium leprae. The clinical findings from tuberculoid to lepromatous leprosy are a result of variation in the cellular immune response to the mycobacterium. The resulting impairment of nerve function causes the disabilities associated with leprosy. The widespread implementation of multi-drug therapy (MDT) has been associated with a fall in the prevalence of the leprosy but as yet no reduction in the case-detection rate globally. The leprosy control activities must be maintained for decades to interrupt transmission of infection. Based on the discovery of three single nucleotide polymorphisms (SNPs) in Mycobacterium leprae, it has been previously reported that there are four major SNP types associated with different geographic regions around the world. To expand the analysis of geographic distribution of M. leprae, classified by SNP, the author studied 56 clinical isolates from Korea isolates. And analysis SNP genotyping by PCR amplification and sequencing, PCR-RFLP, and pyrosequencing. The genotype of single-nucleotide polymorphism type 3, CTC, at positions 14676, 164275, and 2935685, was predominant (95%) for isolates originating in Korea.
Genotype ; Immunity, Cellular ; Korea ; Leprosy ; Leprosy, Lepromatous ; Mycobacterium ; Mycobacterium leprae ; Peripheral Nerves ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Prevalence ; Skin

BACKGROUND: Polymerase chain reaction(PCR) has brought an oppotunity for rapid detection of Mycobacterium leprae in clinical pecimens for the diagnosis of leprosy. Th DNA segment specific to M. leprae was detectable in a matteir of hours and DNA from one orgnisa appeared positive by PCR. However, the PCR tool has not been evaluated using elinical specimeriis from leprosy patients and controls. OBJECTIVE & METHODS: The primers amplifying 372bp segment of rebetitive sequence of M. leprae DNA were used in PCR. Skin biopsy specimens from 102 leprosy patient, and controls were examined for the presence of M. leprae by PCR and the results were aomared with microscopic and histopathologic findings. RESULTS: 1. As a result, of PCR after DNA preparation of M. leprae, six other mycobacteria, ten other bacteria, and skin from leprosy with five other skin biopsy tissues, 372bp DNA fragment was specifically amplified from M. leprae. 2. Dot blot, hybridization of PCR products showed that the 372bp DNA from skin biopsy specimens were derived from M. leprae. 3. As a result of PCR after DNA preparation of 10-fold diluted M. legrae from mouse footpad, PCR gave a positive result as low as one organism. 4. Of 87 specimens in which acid-fast bacilli were found under microcopic examinations 97% had positive PCR results. 5. Of 97 specimens which hadihistopathologic evidences of leprosy 95% had positive PCR results. 6. Of 15 specimens in which acid-fast bacilli were not found under n!icroscopic examinations 73% had positive PCR results. In three of five cases which had neither histopathologic nor microscopic evidences of leprosy had positive PCR results. CONCLUSION: PCR method amplifying 372bp fragment of repetitive seqi,ence was highly sensitive and specific in detecting M. leprae DNA in skin biopsy specimens, thus may be a useful tool as an additive diagnostic method, espcially for cases where microscopic antihystopathologic findings are not definite.
Animals ; Bacteria ; Biopsy* ; Diagnosis ; DNA ; Humans ; Leprosy* ; Mice ; Mycobacterium leprae* ; Mycobacterium* ; Polymerase Chain Reaction* ; Skin*

According to the reports, the prevalence of anti-HCV is about 3%. In past, the results of the high prevalence of anti-HCV in the patients of Hansen's disease were reported. So we study about the Status of Hepatitis C of persons affected Hansen's disease. The prevalence of anti-HCV is 16%(persons affected Hansen's disease: total), 23%(live in settlement village) and 4.1%(live in home). Positivity of RT PCR is 37.8% in HCV-Ab-positive persons affected Hansen's disease. Statistical signification in age and duration of illness of Hansen's disesae between HCV-Ab-positive cases & HCV-Ab-positive case is found (P<0.01, P<0.01).
Hepacivirus ; Hepatitis C* ; Hepatitis* ; Humans ; Jeollabuk-do* ; Leprosy* ; Polymerase Chain Reaction ; Prevalence

According to the reports, the prevalence of anti-HCV is about 1%. In past, the results of the high prevalence of anti-HCV in the patients of Hansen's disease were reported. So we study about the status of Hepatitis C of persons affected Hansen's disease. 1. The prevalence of anti-HCV is 35.1%(persons affected Hansen's disease), 0%(health contact), 0.71%(control, general populations). 2. Positivity of RT PCR is 88% in HCV-Ab-positive persons affected Hansen's disease. In the genotype of hepatitis C, type 1 genotype is 68%, and type 2 is 32%. 3. Statistical signification between patients & non-patients in ALT and HCV-Ab-positivity is found P=0.02, P=0.00) and statistical signification according to result of Hepatitis C antibody in ALT, GTT, and AFP is found in persons affected Hansen's disease(P=0.015, P=0.036, P=0.017).
Genotype ; Hepacivirus ; Hepatitis ; Hepatitis C ; Humans ; Leprosy ; Polymerase Chain Reaction ; Prevalence

Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method that amplifies DNA with high specificity, efficiency, and speed under isothermal conditions. To evaluate the usefulness of LAMP for diagnosing leprosy, we compared the results of LAMP method and PCR, using samples that were previously tested by PCR. We used the species-specific primers designed by targeting the 18KDa antigenic protein gene for the LAMP and PCR, for detection of Mycobacterium leprae directly from biopsy samples of patients. Doing LAMP and PCR, when the results of PCR ware regarded as standard, we observed the results of LAMP were equal to those of PCR. So we are thought the LAMP method is rapid, taking only 1 h, compared with about 4 h for PCR. In addition, it is performed under isothermal conditions and no special apparatus is needed, which makes it more economical and practical than PCR. Although further improvement is necessary for the wide spread use, the LAMP method might be applicable to diagnosis of leprosy.
Biopsy ; Diagnosis* ; DNA ; Humans ; Leprosy* ; Mycobacterium leprae* ; Mycobacterium* ; Polymerase Chain Reaction* ; Sensitivity and Specificity

BACKGROUND: PCR from paraffin-embedded tissues has been recently applied on the diagnosis of leprosy, PCR is a highly sensitive technique and the amplified product is detected by gel electrophoresis and Southern blot hybridization. But conventional PCR does not give an information about histopathologic features. On the other hand in situ PCR informs the histopathological location of DNA amplified inside the cells and detected by in sita hybridization or immunohistoche mical method. OBJECTIVE: In situ PCR was applied on the paraffin-embedded tissues of borderline leprosy patients. The optimal condition of in situ PCR in leprosy was searched with the parameters of pretreatment of the tissues, fixation time of paraformaldehyde and total volume of PCR. METHODS: The paraffin-embedded tissues of ten borderline leprosy patients were subjected to in situ PCR. Amplified DNA product within the cells was incorporated with Digoxigenin and was directly detected by immunohistochemical method using anti-Dig-alkaline phosphatase conjugated antibody. The tissues were pretreated with 0.2N HCl or various concentration of proteinase K such as 10, 15 and 100ug/ml and various incubation time of proteinase K from 3.5 minutes to 15 minutes. Fixation was performed before pretreatment. and after PCR for 15 minutes, after pretreatment and after PCR for 15 minutes, only after pretreatment for 15 minutes or after pretretmant for 60 minutes. The total volume of PCR was 40, 50 or 60ul. RESULTS: 1. The amplified DNA was detected using the HCl-pretreated tissues in four of seven BL patients and one of three BT pat ients. 2. The signals were detected in the cytoplasm of most histiocytes and proliferated Schwann cells, some secretory cells of sweat glands and a few endothelial cells in the tissues of the BL patients and the cells composing of the granuloma in those of the BT patients. 3. Proteinase K-treated tissues showed positive reaction in only one tissue used in 10ug/ml of proteinase K for 10 minutes. 4. The proper time for fixation of paraformaldehyde was before treatment of proteinase K or HCl and after PCR reaction. 5. The 40ul of total volume for PCR reaction was enough and minimized the loss of tissues during PCR reaction. CONCLUSION: When in situ PCR was applied on the paraffin-ernbedded tissues of borderline leprosy patients, pretreatment such as concentration and incubation time of proteinase K or HCL was the most critical parameter.
Blotting, Southern ; Cytoplasm ; Diagnosis ; Digoxigenin ; DNA ; Electrophoresis ; Endopeptidase K ; Endothelial Cells ; Granuloma ; Hand ; Histiocytes ; Humans ; Leprosy ; Leprosy, Borderline* ; Polymerase Chain Reaction* ; Schwann Cells ; Sweat Glands