Pure neuritic leprosy in patients from a high endemic region of Colombia.

Agua de Dios was a leprosarium for leprosy patients' obligatory isolation (1872-1961). Its leprosy incidence is the highest in Colombia (1.5-7/10000). Relapses are common. Government grant of US$ 200 per month subsidy is available to patients with disabilities. Spontaneous consultation with neural symptoms is frequent and simulation to get the subsidy has to be considered. We studied 36 subjects (2007-2009), with ages from 29-78, 19 of them men, with neural symptoms of 6 months to 20 years evolution. All had clinical examination, bacteriological examination, skin and nerve biopsies, electromyography (EMG), PCR for M. leprae, IgM anti-PGL1, and lepromin A. All but two are household contacts of leprosy patients. Symptoms were hypoesthesia of the hands and feet, and difficulty using hands with loss of muscular strength. None had skin lesions. Three had thickening of ulnar nerve. Lepromin was positive in all; bacteriology and biopsies were negative in all. The speed and amplitude of neural conduction were altered in 34 patients; two women had normal EMG and were considered to be feigning the disease; 21 were diagnosed as PNL by clinical, epidemiological and EMG findings; five of them had a positive PCR and one, high titers for IgM anti PGL1. Nine other subjects had diabetes and six carpal tunnel syndrome (CTS). Slow progression of disease, the lack of neural enlargement and the neural biopsies without inflammation suggest that most of these patients could have spontaneously cured PNL, as happens with other cases of paucibacillary leprosy. Diabetes and CTS are important differential diagnoses of PNL. Patients were treated with MDT and received the state subsidy.

Identification and comparison of Mycobacterium leprae genotypes in two geographical regions of Colombia.

OBJECTIVE: To evaluate and establish genomic strain typing markers suitable for the identification of transmission patterns of leprosy in different regions of Colombia. DESIGN: Patients from Agua de Dios, Barranquilla and Cartagena cities and neighbouring towns were enrolled during 2006-2007. Slit skin smears or biopsies were obtained from newly detected untreated patients, and those undergoing multidrug therapy. DNA was extracted from the clinical samples and tested using 15 different short tandem repeat and three SNP polymorphic markers. RESULTS AND CONCLUSION: Differences or similarities between strain types from the northeast (n = 20) and central regions of Colombia (n = 18) were noted. The alleles at two loci, 27-5 and 12-5 were different in the M. leprae in the two regions. The other microsatellite loci may be useful for further intra-population differentiation. There was strong association of 27-5 and 12-5 alleles with the SNP types. The 4-5 combination of alleles was associated with SNP type 3, while the 5-4 combination was mostly associated with SNP type 1, 2 or 4. The SNP type 4 m. leprae isolates were seen in patients in the northeast, but not in the central part.

Clinical, bacteriological and immunological follow-up of household contacts of leprosy patients from a post-elimination area - Antioquia, Colombia.

Follow-up of the household contacts (HHC) of leprosy patients is still the best strategy for early detection of leprosy. HHC from a post-elimination region of Colombia studied in 2001-2002 were re-contacted in 2007. They were tested at both times by clinical examination, bacillary index (BI), PCR from a slit skin smear (SSS) and anti PGL-1 IgM titres. Thirty-two of 61 HHC (52%) were re-contacted. Nine HHC (28%) showed sero-conversion and one had a skin lesion (BI negative, nested PCR positive). Periodic evaluation of HHC can contribute to the detection of infected HHC as well as new and early leprosy cases.

Comparison of Enzyme-Linked Immunosorbent Assay Using Either Natural Octyl Disaccharide-Leprosy IDRI Diagnostic or Phenolic Glycolipid-I Antigens for the Detection of Leprosy Patients in Colombia.

Leprosy is a chronic infectious disease with a broad spectrum of manifestations. Delays in attaining correct diagnosis permit progressive peripheral nerve damage that can produce irreversible disabilities. Tests detecting antigen-specific antibodies can aid the diagnostic process and potentially detect patients earlier. Reported tests have lacked optimal sensitivity and specificity; however, the need to develop new tests to aid early diagnosis still remains. In this study, we determined the sensitivity, specificity, positive predictive value, and negative predictive value of enzyme-linked immunosorbent assay (ELISA) using natural octyl disaccharide-leprosy IDRI diagnostic (NDO-LID). Serum samples from confirmed multibacillary patients (N = 338) and paucibacillary patients (N = 58) were evaluated and contrasted against samples from individuals without leprosy (100 healthy persons, 36 leishmaniasis or tuberculosis patients). ELISA detecting either antigen-specific IgM, IgG, or the combination of IgG and IgM (with protein A) were conducted. At a sensitivity of 78% among all patients, serum IgM antibodies against the NDO-LID conjugate were detected at a greater level than those recognizing phenolic glycolipid-I antigen (64% overall sensitivity), while providing similar specificity (97% versus 100%, respectively). Given the inclusion of the LID-1 protein within NDO-LID, we also detected conjugate-specific IgG within patient sera at a sensitivity of 81.6%. The use of protein A to simultaneously detect both antigen-specific IgG and IgM isotypes yielded the highest overall sensitivity of 86.3%. Taken together, our data indicate that the detection of both IgG and IgM antibodies against NDO-LID with protein A provided the best overall ability to detect Colombian leprosy patients.

Evaluation and Monitoring of Mycobacterium leprae Transmission in Household Contacts of Patients with Hansen's Disease in Colombia.

Leprosy in Colombia is in a stage of post elimination-since 1997, prevalence of the disease is less than 1/10000. However, the incidence of leprosy has remained stable, with 400-500 new cases reported annually, with MB leprosy representing 70% of these case and 10% having grade 2 disability. Thus, leprosy transmission is still occurring, and household contacts (HHCs) of leprosy patients are a population at high risk of contracting and suffering from the effects of the disease during their lifetime. We performed a cross-sectional study with the aim of evaluating leprosy transmission within Family Groups (FGs) from four Colombian departments: Antioquia, Bolívar, Córdoba and Sucre. This study included 159 FGs formed by 543 HHCs; 45 FGs were monitored twice, first in 2003 and again in 2012. Migration, forced displacement by violence, loss of contact with the health center and the lack of an agreement to participate in the second monitoring were the primary reasons not all FGs were tested a second time. In each HHC, a clinical examination was performed, epidemiological data recorded, the bacillary index determined, DNA was isolated for M. leprae detection by nested PCR and IgM anti-phenolic glycolipid-I (PGL-I) titers were inspected. Further, DNA from M. leprae isolates were typed and compared among FGs. Twenty-two (4.1%) of the 543 HHCs had IgM anti-PGL-I positive antibody titers, indicating infection. Nasal swabs (NS) taken from 113 HHCs were tested by RLEP PCR; 18 (16%) were positive for M. leprae DNA and two new leprosy cases were detected among the HHCs. Of the confirmed HHCs with leprosy, it was possible to genotype the bacterial strains from both the index case and their HHCs. We found that the genotype of these two strains agreed at 9 markers, showing the individuals to be infected by the same strain, indicating familiar transmission. HHCs of leprosy patients not only are a high-risk population for M. leprae infection, they can act as M. leprae carriers and therefore serve as sources for transmission and infection. Our results confirm familiar leprosy transmission and suggest that follow-up of HHCs is a good strategy for early diagnosis of leprosy and to monitor its transmission.

Leprosy in Colombian children and adolescents.

While leprosy is not considered a public health problem in Colombia, affected children are an important warning sign, demonstrating the challenge of controlling the disease. Herein, we report 12 cases of leprosy in patients 5-17 years of age, summarizing our clinical, micro biological and treatment findings.

A rapid ELISA for the diagnosis of MB leprosy based on complementary detection of antibodies against a novel protein-glycolipid conjugate.

Despite the widespread use of multidrug therapy for treatment, delays in clinical recognition and under-reporting of leprosy indicate that Mycobacterium leprae transmission is continuing. Thus, leprosy is likely to persist as a significant burden on health systems in many regions. In this study, we combined 2 previously characterized leprosy antigens, leprosy IDRI diagnostic-1 (LID-1) and ND-O, into the single fusion complex (ND-O-LID) and determined the serum antibody responses of leprosy patients from Colombia and the Philippines. Following confirmation that antibodies recognized each component within the conjugate, we assessed the performance of a rapid enzyme-linked immunosorbent assay (ELISA) system (Leprosy Detect(TM) fast ELISA; InBios International, Inc., Seattle, WA, USA) based on ND-O-LID capable of generating results within 1.5hours of sample addition. We found ELISA results correlated with the bacteriological index and Ridley-Jopling categorization, with lepromatous leprosy patients having the highest responses, while those of borderline tuberculoid patients were lower. Multibacillary (MB) leprosy patients were distinguished with a high degree of sensitivity (95.7%) and specificity (93.2%), suggesting that this ELISA could potentially replace invasive and insensitive skin slit smear procedures that require expert microscopic examinations. Due to the speed and robustness of this assay, we believe this is an excellent tool for detecting MB leprosy patients in a simple and highly-quantitative manner.

Mycobacterium leprae in Colombia described by SNP7614 in gyrA, two minisatellites and geography.

New cases of leprosy are still being detected in Colombia after the country declared achievement of the WHO defined 'elimination' status. To study the ecology of leprosy in endemic regions, a combination of geographic and molecular tools were applied for a group of 201 multibacillary patients including six multi-case families from eleven departments. The location (latitude and longitude) of patient residences were mapped. Slit skin smears and/or skin biopsies were collected and DNA was extracted. Standard agarose gel electrophoresis following a multiplex PCR-was developed for rapid and inexpensive strain typing of Mycobacterium leprae based on copy numbers of two VNTR minisatellite loci 27-5 and 12-5. A SNP (C/T) in gyrA (SNP7614) was mapped by introducing a novel PCR-RFLP into an ongoing drug resistance surveillance effort. Multiple genotypes were detected combining the three molecular markers. The two frequent genotypes in Colombia were SNP7614(C)/27-5(5)/12-5(4) [C54] predominantly distributed in the Atlantic departments and SNP7614 (T)/27-5(4)/12-5(5) [T45] associated with the Andean departments. A novel genotype SNP7614 (C)/27-5(6)/12-5(4) [C64] was detected in cities along the Magdalena river which separates the Andean from Atlantic departments; a subset was further characterized showing association with a rare allele of minisatellite 23-3 and the SNP type 1 of M. leprae. The genotypes within intra-family cases were conserved. Overall, this is the first large scale study that utilized simple and rapid assay formats for identification of major strain types and their distribution in Colombia. It provides the framework for further strain type discrimination and geographic information systems as tools for tracing transmission of leprosy.

Lepromatous leprosy and human immunodeficiency virus co-infection associated with phenomenon of Lucio versus immune reconstitution inflammatory syndrome / Lepra lepromatosa y coinfección con el virus de la inmunodeficiencia humana asociada a fenómeno de Lucio versus síndrome inflamatorio de reconstitución inmune

Diffuse lepromatous leprosy (DLL) is a severe clinical outcome of lepromatous leprosy (LL). The aetiologic cause is believed to be different from Mycobacterium leprae. A new species, Mycobacterium lepromatosis, was identified from a group of Mexican patients with DLL, and severe leprosy reactional state type 3 (Lucio's phenomenon). However, a total sequencing of its genome is necessary to prove the existence of this new species. This is a report on a non-typical Colombian case of leprosy - HIV coinfection, associated with an immune reconstitution inflammatory syndrome clinically compatible with a leprosy reaction type 3 or Lucio's phenomenon.

La lepra difusa (LLD) es una variedad de la lepra lepromatosa (LL), frecuente enMéxico. El agente etiológico se cree que es diferente a Mycobacterium leprae y se considerauna especie nueva denominada Mycobacterium lepromatosis, hecho que no se ha comprobado.El reporte de este caso se realiza para dar a conocer el cuadro clínico atípico que presentóuna paciente colombiana con coinfección VIH---LL variedad difusa (LLD), asociado a síndromede reconstitución inmunológica, compatible clínicamente con una leprorreacción tipo 3 o fenó-meno de Lucio.

Clinical, bacteriological and immunological follow-up of household contacts of leprosy patients from a post-elimination area - Antioquia, Colombia

Follow-up of the household contacts (HHC) of leprosy patients is still the best strategy for early detection of leprosy. HHC from a post-elimination region of Colombia studied in 2001-2002 were re-contacted in 2007. They were tested at both times by clinical examination, bacillary index (BI), PCR from a slit skin smear (SSS) and anti PGL-1 IgM titres. Thirty-two of 61 HHC (52 percent) were re-contacted. Nine HHC (28 percent) showed sero-conversion and one had a skin lesion (BI negative, nested PCR positive). Periodic evaluation of HHC can contribute to the detection of infected HHC as well as new and early leprosy cases.

Búsqueda de úlcera de Buruli en el Urabá Chocoano y Antioqueño, Colombia 2006 / Search of Buruli in Choco and Antioquia, Uraba Colombia, 2006

Introducción: en Colombia no se ha reportado ningún caso de úlcera de Buruli (UB), aún teniendo regiones con características similares a zonas endémicas. En nuestro medio, la proximidad geográfica y las condiciones ambientales similares con los países de Sur y Centro América donde se han reportado casos, motivan a buscar activamente pacientes sospechosos de UB, y aplicar técnicas de laboratorio moleculares específicas para brindar un adecuado diagnóstico. Objetivo: buscar casos de úlcera de Buruli (UB) en Urabá chocoano y antioqueño, (Colombia) durante el año 2006. Materiales y métodos: se estudiaron casos provenientes de las áreas de estudio, para establecer la causa etiológica de las lesiones utilizando métodos de diagnóstico clínico, microbiológico, histopatológico y molecular. Resultados: en cinco pacientes (6%) no se pudo establecer la causa etiológica de la úlcera (leishmaniosis, micosis, úlceras venosas o arteriales, cáncer). El examen clínico de estos pacientes no fue concluyente de UB, sin embargo debido a la falta de documentación de casos en Colombia, se procesó biopsia de la lesión para detectar ADN de Mycobacterium ulcerans por reacción en cadena de la polimerasa (PCR). Todas las pruebas de PCR fueron negativas para ADN de Mycobacterium ulcerans. Conclusiones: debido a indicadores epidemiológicos que señalan a Colombia con condiciones geográficas y ambientales similares a las que se presentan en regiones endémicas, como Perú, Guyana Francesa, México, Surinam, es necesario continuar con su búsqueda.

Introduction: in Colombia there are no reported cases of Buruli ulcer (BU), however the geographic and environmental characteristics are similar to endemic regions, and the proximity to other countries in South and Central America where there are reported cases, makes it an important issue to search for them, using molecular techniques specific for BU diagnostic. Objectives: to search for Buruli ulcer (BU) at the Urabá region of Chocó and Antioquia in Colombia, during 2006. Materials and methods: patients with skin ulcer from the study region were tested to establish the etiologic cause of the lesions, using clinical, microbiological, pathological and molecular methods. Results: Five patients were tested for BU using PCR test, since other etiologic causes of the ulcer (leishmaniosis, mycosis, venous or arterial ulcer, others) were not determined. Clinical examination of the patients was not conclusive of BU; but due to the lack of documented cases in Colombia, biopsies were taken from patients for detection of M. ulcerans by PCR. All the samples samples tested negative for DNA of Mycobacterium ulcerans. Conclusions: due to the epidemiological indicators that show that Colombia has the geographic and environmental conditions similar to endemic regions, as Perú, French Guyana, México and Surinam it is necessary to continue with the search.