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*

Hansen's disease(HD) is a chronic infectious disorder acquired by inoculation of Mycobacterium leprae. With the establishment of complex multidrug therapy, the incidence rate of leprosy patients has continually shown to decline by 90% compared to the incidence rate in the 1990s. However, the prevalence of the disease still remains high in southeast asian countries. Due to the rarity and diverse nature of cutaneous presentation, HD is often misdiagnosed with other dermatoses or infectious conditions. Especially, when a patient presents with unusual presentation with leprosy reaction with no classical feature such as sensory disorders and skin lesion, the diagnosis is further delayed with misguided treatments. Herein we present a 27-year-old Indonesian immigrant who displayed clinical features mimicking that of orbital cellulitis who was later diagnosed with borderline lepromatous leprosy through histologic and PCR confirmation, in light of alerting the probability of leprosy in immigrants with intractable skin presentations.
Adult ; Asian Continental Ancestry Group ; Diagnosis ; Emigrants and Immigrants ; Humans ; Incidence ; Leprosy ; Leprosy, Borderline ; Leprosy, Multibacillary ; Mycobacterium leprae ; Orbit ; Orbital Cellulitis ; Polymerase Chain Reaction ; Prevalence ; Sensation Disorders ; Skin ; Skin Diseases

Adult ; Female ; Heat-Shock Proteins ; genetics ; Humans ; Leprosy ; diagnosis ; genetics ; microbiology ; Male ; Middle Aged ; Mycobacterium leprae ; pathogenicity ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Skin ; metabolism

BACKGROUND: Mycobacterium leprae, the etiological agent of leprosy, is noncultivable on axenic media, so, the viability of M. leprae for clinical or experimental applications is often unknown. Quantitative reverse transcriptase PCR (RT-PCR) assays were recently introduced as the new tools for M. leprae viability determination. OBJECTIVE: For evaluating of correlation of results of quantitative real-time PCR(16S rRNA/RLEP) & AFB stain of slit skin smear & histopathology & estimating the viability of M. leprae, the author studied the comparison of results of them METHODS: Of 46 samples from 27 patients(MB 24 cases, PB 3 cases), M. leprae 16S rRNA was used as RNA targets, and M. leprae repetitive element (RLEP) DNA was used to determine relative bacterial numbers and the viability by the quantitative real-time PCR. The ratio of 16S rRNA and RLEP as the indicator of viability was calculated. Student t test and linear Pearson correlation were done by SPSS. RESULTS: There was a correlation between between 16S rRNA/RLEP ratio and BI (r=0.369, p=0.012), and was statistically significant between 16S rRNA/RLEP ratio and histopathological positivity of AFB (p=0.011). However there was no correlation between 16S rRNA/RLEP ratio and MI. CONCLUSIONS: Although the correlation between between 16S rRNA/RLEP ratio and BI and the statistically significant between 16S rRNA/RLEP ratio and histopathological positivity of AFB, there was no correlation between 16S rRNA/RLEP ratio and MI. It needs the further evaluation the correlation about that.
DNA ; Humans ; Leprosy* ; Mycobacterium leprae ; Real-Time Polymerase Chain Reaction* ; Reverse Transcriptase Polymerase Chain Reaction ; RNA ; 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

The Ziehl-Neelson's AFB staining method was mainly used for the AFB observation of the diagnosis of leprosy. However, the fluorescent stain performs better and allows the detection of more positive smears. The limitation for its widespread use has been the high cost for fluorescent microscopes. Novel light-emitting diodes (LED) are inexpensive solutions for fluorescent microscopes, and thus fluorescent stain may be a cost-effective step to improve the diagnosis of leprosy in resource-poor countries. And the comparison of auramine and acridine orange for staining of acid-fast bacteria was showed significantly more acid-fast rods after using acridine orange and the number of "false positive" results was somewhat higher on auramine staining. So acridine orange offers a good alternative to auramine which is considered carcinogenic. This study evaluated the comparison of the Ziehl-Neelson's AFB stain and the acridine orange stain in the skin smear based on PCR. As PCR results were taken as gold standard, results of the study revealed that the sensitivity of Ziehl-Neelson's AFB stain was 50% and that of acridine orange stain was 92.2%. This study confirmed that the fluorescence stain method is more sensitive than the Ziehl-Neelsen's staining method. It is suggested that the training of laboratory technicians on fluorescence microscopy should be scaled up for increased disease control.
Acridine Orange* ; Bacteria ; Benzophenoneidum ; Diagnosis ; Fluorescence ; Humans ; Laboratory Personnel ; Leprosy ; Microscopy, Fluorescence ; Mycobacterium leprae* ; Mycobacterium* ; Polymerase Chain Reaction ; Skin

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

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

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

OBJECTIVEMultiple locus variable number-tandem repeat (VNTR) analysis (MLVA) had been proposed as a means of strain typing for tracking of source and studying the transmission chain of pathogens. However, empirical data for a defined population from scale and duration were lacking for studying the transmission chain of leprosy.

METHODSMLVA on 7 VNTR loci was applied to the strain typing on prevalent Mycobacterium leprae isolates collected from Qiubei county, Yunnan province during 2002-2006 in the study on the relationship between geographic distribution and genotypes of M. leprae. The strain typing, combined with conventional epidemiological investigation was performed to trace the transmission chain.

RESULTS(1) Phylogenetic analyses through application of PAUP 4.0, The M. leprae were grouped into A, B, C, D and E strains according to the allelic range 9, 11-13, 15-26 and > 26 on the GTA9 locus. The strains with 9 copies on GTA9 locus, was named A. (2) Genotypes of strains from the five multi-case families located at North and North-West parts were similar and belonged to A strains. VNTR patterns of intra-family were identical or similar but not identical inter-family. (3) Not only A cluster appeared higher proportion in total isolates but also distributes cluster, indicating ongoing transmission from recent findings.

CONCLUSIONVNTR strain typing was suitable to trace the short chain of transmission in both small area and intra-families. Multi-case families might constitute epidemic foci and source of M. leprae in villages, causing the predominant strain or cluster which tends to be those identified in multi-case families and resulted in the spreading of leprosy. A long-term study was underway to reveal whether A strain was predominant strain and to observe the evolution of M. leprae in this spatially and temporally defined endemic population.

Female ; Genotype ; Humans ; Leprosy ; microbiology ; Male ; Minisatellite Repeats ; genetics ; Molecular Epidemiology ; Mycobacterium leprae ; classification ; genetics ; Phylogeny ; Polymerase Chain Reaction