Identification of trehalose dimycolate (cord factor) in Mycobacterium leprae

AbstractGlycolipids of Mycobacterium leprae obtained from armadillo tissue nodules infected with the bacteria were analyzed. Mass spectrometric analysis of the glycolipids indicated the presence of trehalose 6,6′-dimycolate (TDM) together with trehalose 6-monomycolate (TMM) and phenolic glycolipid-I (PGL-I). The analysis showed that M. leprae-derived TDM and TMM possessed both α- and keto-mycolates centering at C78 in the former and at C81 or 83 in the latter subclasses, respectively. For the first time, MALDI-TOF mass analyses showed the presence of TDM in M. leprae

Genotyping Analysis of Mycobacterium leprae isolated in Water Environment of Leprosy Endemic Places in Lamongan, East Java

Finding of Mycobacterium leprae from water of leprosy endemic areas were reported. East Java Province is ranked number eight as the endemic area of leprosy in Indonesia and Lamongan district is the local area with the highest new cases reported. To study the transmission of Mycobacterium leprae infection in endemic areas, it is important to detect the distribution of Mycobacterium leprae in the environment and population, also to analyze the genetic variation pattern. Keywords: endemic; environment; genotyping; leprosy; Mycobacterium leprae

Variation of TTC Repeat Pattern in the Dna of Mycobacterium Leprae Isolates Obtained From Archeological Bones and Leprosy Patients From East Nusa Tenggara

The existence of leprosy or kusta or Morbus Hansen or Hansens disease has been known for years, including in Indonesia. Starting from the discovery of Mycobacterium leprae isolates from ancient bone (about 1.000 years B.C), the archaeological excavations results in East Nusa Tenggara, interesting questions arise about how the development of leprosy in eastern Indonesia is. Biology molecular study would become a powerful tool to investigate the presence of leprosy bacillary whether there are similarities between the genomes of M. leprae isolates in the primeval and the present. PCR examinations were performed on mandibular bone fragments from ancient human who lived 1000 years B.C. discovered in archaeological surveys on the island of Lembata and three leprosy patients from East Nusa Tenggara. The DNA extraction was performed using a kit from Qiagen products and its TTC repeating pattern was seen with the method of direct sequencing. It turned out that the TTC profile obtained from samples of archaeological was as many as 13 copies, while the repetition of TTC in three samples of leprosy patients were 15, 17 and 26 copies. The different number of TTC repetition shows the different isolates of M. leprae between in the ancient times and the present. Further studies are needed to verify the differences in the genome that occur, for example from the study of SNPs (single nucleotide polymorphisms)

Potential for serodiagnosis of indonesian leprosy patients by detecting antibodies against LID-1

Leprosy is caused by Mycobacterium leprae infection and remains a major public health problem in many areas of the world. The Leprosy IDRI Diagnostic (LID)-1 antigen has demonstrated potential to improve the diagnostic capacity of local health centers and aid the development of strategies for the eventual control and elimination of leprosy from endemic areas. The diagnostic capacity of LID-1 has not yet, however, been studied in Indonesia. Objective: To determine the proportion of Indonesian leprosy patients that have circulating antibodies against LID-1. Sera from thirtyfive leprosy patients from Surabaya, Indonesia were evaluated using an Enzyme Linked Immunosorbent Assay (ELISA) detecting IgG antibodies against the LID-1 antigen. Anti-LID-1 antibody levels correlated with both the clinical form of leprosy and the bacterial index (BI). LID-1-specific IgG responses were higher in multibacillary (MB) than in paucibacillary (PB) leprosy patients. Our data indicate that the detection and measurement of serum IgG against LID-1 could be an effective tool for use in control programs in various states and municipalities in Indonesia

ACANTHAMOEBA SP.S-11 PHAGOCYTOTIC ACTIVITY ON MYCOBACTERIUM LEPRAE IN DIFFERENT NUTRIENT CONDITIONS

Background: Mycobacterium leprae (M. leprae) is a pathogenic bacterium that causes leprosy. The presence of M. leprae in the environment is supported by microorganisms that act as the new host for M. leprae. Acanthamoeba’s potential to be a host of M. leprae in the environment. Acanthamoeba sp. is Free Living Amoeba (FLA) that classified as holozoic, saprophytic, and saprozoic. The existence of nutrients in the environment influence Acanthamoeba ability to phagocytosis or pinocytosis. This study is aimed to determine Acanthamoeba sp.S-11 phagocytic activity to Mycobacterium leprae (M. leprae) which cultured in non-nutrient media and riched-nutrient media. Materials and Methods: This research conducted by culturing Acanthamoeba sp.S-11 and M. leprae on different nutrient media conditions. M. leprae intracellular DNA were isolated and amplified by M. leprae specific primers through Real Time PCR (Q-PCR). Result: The results showed that Acanthamoeba co-cultured on non-nutrient media were more active to phagocyte M. leprae than on rich-nutrient media. Conclusion: The use of non-nutrient media is recommended to optimize Acanthamoeba sp. phagocytic activity to M. leprae. Keywords: Acanthamoeba sp., Mycbacterium leprae, Nutrient, Phagocytosis, Real Time PCR Introduction Mycobacterium leprae (M. leprae) is a pathogeni

The Correlation of Ig M Anti PGL-1 antibody between blood veins and dryed capillary blood on filter papers in household contact of leprosy patient

Delays of leprosy detection and treatment can lead to disability and potential transmission. Serologic examination has the advantage in detecting Subclinical Leprosy. The procedure of serologic test, which is one of its main limitation, could be simplified by the use filter paper. This study aims to assess the effectiveness of the use of capillary blood dropped on filter paper as a substitute for venous blood in household contact of leprosy patients. Seventeen samples of capillary blood dried on filter paper and venous blood samples from the same individual were examined by ELISA method to determine the levels of IgM anti-Phenolic glycolipid-1 (PGL-1). The mean of anti- PGL-1 IgM levels of filter paper samples 163.31±126.16; whereas the mean of levels from venous samples was 473.16±411.26. There was significant difference and correlation between these two groups. Samples on filter paper in household contact can be used to determine the level of anti-PGL-1 IgM in serum by converting on the regression basis. Further study is required to evaluate the potency of filter paper methods to conduct large-scale serological screening

Distribution of Mycobacterium leprae genotypes from Surabaya and Bandung Clinical Isolates by Multiple Locus Variable Number of Tandem Repeat Analysis

Multiple Locus Variable Number of Tandem Repeat (VNTR) analysis has been proposed as a means of genotyping for tracking leprosy transmission due to tandem repeats’ potential as genetic markers to differentiate M. leprae strains. However, characteristics of polymorphism can vary depending on the population. This study aimed to compare the copy number of repeats in four genetic markers: TTC, AC8a, AC9 and 6-7 in leprosy patients from Surabaya and Bandung. Twenty three patients from Dr. Soetomo General Hospital and 21 from Hasan Sadikin Hospital were recruited. Multiple locus VNTR analysis was applied using total DNA extracts from Slit Skin Smear (SSS). From Surabaya, 7 samples showed the same copy number of four genetic markers (TTC=15; AC8a=10; AC9=10 and 6-7=6) and 2 showed another (TTC=16; AC8a=10; AC9=11 and 6-7=6); as for samples from Bandung, 2 showed the same copy number (TTC=15; AC8a=8; AC9=10 and 6-7=8) and 2 showed another (TTC=16; AC8a=10; AC9=11 and 6-7=6). The multiple locus VNTR analysis showed two identical M. leprae VNTR profiles from Bandung and Surabaya which supports the use of VNTR loci for transmission studies

A simpler diagnostic method using blood collection on filter paper to determine anti-natural octyl disaccharide-leprosy infectious disease research institute diagnostic in household contacts of leprosy patients

The high prevalence among children shows continued transmission from adult and lack of disease control by the health system. Detection of anti-NDO-LID-1 has been known to be more effective to predict the development of leprosy in household contacts than just detecting PGL-1 or LID- 1 alone. However, serodiagnosis is not available in poorer-settings area and the procedure (venepuncture) is still inconvenient to some people. These problems can be solved by using finger-prick blood sample on filter paper. This study aims to prove the effectiveness of using capillary blood samples on filter paper to detect the positivity of Ig G antibody against NDO-LID-1 antigen in asymptomatic household contacts. Seventeen samples of capillary blood on filter paper and sera were tested for IgG anti-NDO-LID-1 using ELISA. There was no significant difference between IgG level from filter paper and serum (p=0.754) and there was also a strong positive correlation (R=0.906) between the two procedures. These findings show that the use of filter paper and NDO-LID-1 is worthy of further investigations, especially for those with lower bacillary load or contacts of leprosy patients

Immunoglobulin AMG Anti Natural Disaccharide Octyl - Leprosy IDRI Diagnostic (NDO-LID) Serologic Test for Leprosy Diagnosis: a Pilot Study

Leprosy remains a public health problem in Indonesia. Diagnostic tools have been developed in order to aid early diagnosis and prompt treatment. Phenolic glycolipid (PGL)-1 has been considered as a good candidate for leprosy diagnosis but has been found to have several limitations. More recently, the conjugation between natural disaccharide octyl (NDO) and leprosy IDRI diagnostic (LID)-1, known as NDO-LID, shows great promise because of its high specificity and sensitivity and its ability to detect leprosy before any clinical signs are present. The test incorporates the detection of IgM antibodies towards NDO and IgG antibodies towards LID-1. This study aimed to show the profile of IgM, IgG, and IgAMG antibody titers against NDO-LID to further discover its diagnostic potential. Sera from eight new leprosy patients from Surabaya, Indonesia were evaluated using ELISA detecting levels of IgG, IgM and IgAMG antibodies against NDO-LID antigen. Skin slit biopsy was also taken for smear and histopathology test. This study shows that the titer levels IgAMG anti NDO-LID were consistent with the results from smear and were consistently higher compared to IgM or IgG titer alone. IgAMG might have the potential to improve the sensitivity of NDO-LID serologic tests but further investigation is needed

A 5‑year evaluation of chemoprophylactic treatment in elementary school children with subclinical leprosy

Subclinical leprosy is an infectious disease in which the immune system remains infected with Mycobacterium leprae (M. leprae). The progress of subclinical leprosy to clinical cases within 1 year of infection is 1.5%, with an increase to 6% in the following 4 years. Rifampicin is frequently used for prevention of leprosy, and clarithromycin has a bactericidal effect on M. leprae. Thus, the combination of both is expected to improve disease control in patients with subclinical leprosy. The aim of the present study was to evaluate the efficacy of a chemoprophylactic treatment involving rifampicin and clarithromycin against subclinical leprosy in elementary school children from endemic areas of East Java over a 5‑year period. The study was performed between 2011 and 2015. Samples were collected from 2,548 healthy elementary school children in Nguling (Pasuruan) and Raas (Sumenep), and analysed using ELISA for anti‑PGL (phenolic glycolipid)‑1 IgM antibodies. Children who were seropositive for anti‑PGL‑1 IgM antibodies received a chemoprophylactic regimen consisting of rifampicin (300 mg/day) and clarithromycin (250 mg/day) daily for the initial 10 days, followed by the same regimen every 2 weeks for 3 months. Clinical and serological evaluations were performed annually for 5 years. Amongst the 2,548 healthy elementary school children, 200 were seropositive. The anti‑PGL‑1 IgM antibody levels significantly decreased between 2011 and 2015 in Nguling (from 1,066.7 to 137.4 U/ml) and Raas (from 773.1 to 563.4 U/ml), the levels decreased every year. In addition, the proportion of patients with decreased anti‑PGL‑1 IgM antibody levels was consistently higher than patients with increased anti‑PGL‑1 IgM antibody levels in all periods, except during 2013‑2014, in Nguling and Raas. Chemoprophylactic treatment involving rifampicin and clarithromycin may thus be effective against subclinical leprosy amongst elementary school children