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Molecular Detection of Dapsone and Rifampicin Resistance on Mycobacterium leprae from Leprosy Patients in East Java ADRIATY, DINAR; WAHYUNI, RATNA; PRAKOESWA, CITA ROSITA S.; SUSARI, NI PUTU; AGUSNI, INDROPO; IZUMI, SHINZO
Microbiology Indonesia Vol 3, No 3 (2009): December 2009
Publisher : Indonesian Society for microbiology

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Abstract

The drug resistant problem of Mycobacterium leprae has been developing since the last decade and this has become a leprosy elimination problem in several countries, including Indonesia. Using biological on molecular methods, it is now possible to test for drug resistant cases in relatively simple and less time consuming ways. The purpose of the study is to analyze theprevalence of drug resistance M. leprae to dapsone and rifampicin in East Java based on the  etection of mutations in the folP and rpoB genes. All samples were obtained from multibacillary leprosy patients in East Java, who have admitted to the Dr Sutomo Hospital Surabaya in 2003-2005. Isolates were analyzed by PCR, and the presence of nucleotide sequence of the folP and rpoB genes from M. leprae were confirmed by direct sequencing. Of 94 specimens which were collected, all were analyzed for their folP and rpoB genome. From 94 isolates, 70 showed a positive result by the folP1-folPR test and 77 out of 94 isolates showed positive by the rpoBF-rpoBR test. From 70 isolates for folP gene examination, there were 3 isolates which had mutation in the amino acid at codon 53; 2 cases Threonin (ACC) became Alanin (GCC) and 1 case Threonin (ACC) became Arginin (AGA). These mutations are responsible to dapsone resistance. For the rpoB gene, no mutation was found. The result suggested that 3 isolates (4.3%), 1 from a new case and 2 from relapse cases in this experiment, were resistant to dapsone and all isolates (100%) were susceptible to rifampicin.
COMPARATIVE STUDY ON THE INTENSITY OF Mycobacterium leprae EXPOSURE BETWEEN HOUSEHOLD AND NONHOUSEHOLD CONTACT OF LEPROSY Arsyad, Yuniarti; Jifanti, Friska; Amiruddin, Muhammad Dali; Anwar, Anis Irawan; Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi; Agusni, Indropo
Indonesian Journal of Tropical and Infectious Disease Vol 3, No 1 (2012)
Publisher : Institute of Topical Disease

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Abstract

Leprosy stills a public health problem in West Sulawesi which has a Case Detection Rate (CDR) around 43.69/100.000 population. Household contacts of leprosy are a high risk group to be infected, due to droplet infection mode of transmission of the disease. A nose swab examination and serological study was conducted to detect exposure of M. leprae of people who live in leprosy endemic area. Detection of M. leprae in the nasal cavity will represent the exposure rate from outside and the measurement of specific antibody is represented the result of exposure to the immune system. Two group of inhabitants (30 household contacts of leprosy and 30 nonhousehold contacts) were involved in the study. They live in Banggae district, a leprosy endemic area of Majene Regency, West Sulawesi. Sixty nose swab samples and sixty capillary blood samples from the same invidividuals of the two groups were collected and sent to Leprosy laboratory of the Institute of Tropical Disease, Airlangga University Surabaya. A Polymerase Chain Reaction (PCR) was performed to the nose swab samples for detection of M. leprae. The blood samples were examined serologically to measure the level of anti PGL-1 antibody. PCR examination of nose swab samples showed 1/30 positive result in the household contact group and also 1/30 positive result in non-household contact of leprosy (statistically no significant difference, p > 0.05). Serological study showed higher sero-positive result in the household contact group (15/30 or 50%) compared to non-household contact (11/30 or 36%), but statistical calculation revealed no significant difference between the two groups (p > 0.05) on sero-positive results of leprosy. It is concluded that household and non-household contact in leprosy have the same risk to be affected by the disease. The term of household and non-household contact need to be redefined. The possible role of exposure from the environment was also discussed, especially from non-human resource of M. leprae.
COMPARATIVE STUDY ON THE INTENSITY OF Mycobacterium leprae EXPOSURE TO CHILDREN WHO LIVE IN LOW AND HIGH ALTITUDE IN LOW LEPROSY ENDEMIC AREA OF SOUTH SULAWESI Rachmawati, Rachmawati; Mataallo, Timurleng Tonang; Adam, Safruddin; Adam, A. M.; Amin, Safruddin; Tabri, Farida; Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi; Agusni, Indropo
Indonesian Journal of Tropical and Infectious Disease Vol 4, No 4 (2013)
Publisher : Institute of Topical Disease

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Abstract

Background: The intensity of Mycobacterium leprae exposure to people who live in leprosy endemic area could be measured by serological study and detection of the bacilli in the nose cavity. Different geographical altitude might have some influences to this exposure since the bacilli prefer to live in warm areas. Aim: A combined serological and PCR study of leprosy was conducted in Selayar island, South Sulawesi to 80 school children (40 from low land and 40 from highland altitudes) in order to compare the exposure intensity between the two areas. Method: Anti PGL-1 IgM antibody (ELISA) and PCR study to detect M.leprae in the nasal cavity were performed simultaneously from each person. Result: Seropositive cases were found in 23/40 children from low land compared to 16/40 children from high land, but statistically no significant difference (p>0.05). PCR positive for M.leprae in the nasal cavity only found in 1/40 children, both in low and high altitude. Conclusion: It is concluded that although the existence of M.leprae in nasal cavity is minimal, the intensity of exposure to this bacilli still high as indicated by serological study.
Mycobacterium leprae BACILLEMIA IN BOTH TWINS, BUT ONLY MANIFEST AS LEPROSY IN ONE SIBLING Sukmawati, Netty; Agusni, Indropo; Listiawan, M. Yulianto; Prakoeswa, Cita Rosita S.; Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi
Indonesian Journal of Tropical and Infectious Disease Vol 5, No 7 (2015)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/ijtid.v5i7.1206

Abstract

Leprosy in twins is rarely reported. A 19 years-old male student, from Lamongan district, was diagnosed as Multibacillary (MB) leprosy in the Skin and STD Clinic of Dr. Soetomo General Hospital Surabaya. Multiple anesthetic skin lesions were found, but the bacteriologic examination was negative for Acid Fast Bacilli (AFB). Histopathology examination support the diagnosis of BL type of leprosy. His twin brother that has been lived together since born until present seems healthy without any complaints of skin lesions and have no signs of leprosy. When a serologic examination for leprosy was performed, a high anti PGL-1 antibody level was found in patient (IgM anti PGL-1 2937 and IgG anti PGL-1 3080 unit/ml) while his healthy twin brother showed only low level (IgM 745 and IgG 0 unit/ml). Interestingly when a PCR study was performed to detect M.leprae in the blood, both of them showed positive results. Using the TTC method, a genomic study of for M.leprae, it is revealed that both samples were identic ( 27x TTC repeats). According to patient’s history, he had a traffic accident and got a wound in the knee seven years ago, while the skin lesions seems started from this area around three years ago before it spread to other parts of the body. The patient was treated with Multi-drug therapy (MDT) while his sibling got a prophylactic treatment for leprosy. After 6 months of treatment, the leprosy skin lesions were diminished and the serologic anti PGL-1 has been decreased. His healthy brother also showed a decrease in anti PGL-1 level and no skin signs of leprosy.
Mycobacterium leprae in Daily Water Resources of Inhabitants Who Live in Leprosy Endemic Area of East Java Wahyuni, Ratna; Adriaty, Dinar; iswahyudi, iswahyudi; Prakoeswa, Cita Rosita S.; Agusni, Indropo; izumi, shinzo
Indonesian Journal of Tropical and Infectious Disease Vol 1, No 2 (2010)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1204.754 KB) | DOI: 10.20473/ijtid.v1i2.2164

Abstract

Leprosy still a health problem in Indonesia, where many leprosy pocket areas still persists, especially in the eastern part of the country. Although the program of WHO – Multidrug Therapy (MDT) regiment has been conducted elsewhere since 1980s, only the prevalence can be reduced but not the incidence of new leprosy cases. Theoretically after the source of leprosy (the infectious leprosy cases) has been treated, no more transmission of the disease and should be no more new leprosy cases will be found. To explain this phenomenon, the non-human resource of M.leprae became a new topic of debates, especially the existence of bacteria in the environment. A field study of the existence of M.leprae in the environment of leprosy endemic area had been conducted in a leprosy endemic area of the northern part of East Java. The aim of the study is to find any correlation of the existence of these bacteria in the environment with the presence of leprosy patients who live in that area, in order to study its role in the transmission of the disease. Ninety water samples from wells in the house of inhabitants who live in one endemic sub district were collected. The owner of the well was interviewed whether any leprosy patients who routinely use the water for their daily life activities. Water samples were examined by Polymerase Chain Reaction (PCR) method to detect M.leprae DNA, using the LpF-LpR and Lp3-Lp4 nested primers (99bp). The PCR results showed positive band for M.leprae in 22 out of 90 (24%) water samples. Water samples from wells that used by leprosy patients showed positive PCR in 11/48 (23%), while 11 out of 42 (26%) water samples from wells that never been used by leprosy cases showed positive result. Statistically there was no difference (p>0.05) in the positivity of M.leprae between the two groups. It was concluded that the existence of M.leprae in the daily water resource was not correlated with the present of leprosy cases in the area. Possible symbiosis between protozoan and mycobacterium in the environment were discussed.
Mycobacterium leprae BACILLEMIA IN BOTH TWINS, BUT ONLY MANIFEST AS LEPROSY IN ONE SIBLING Sukmawati, Netty; Agusni, Indropo; Listiawan, M. Yulianto; Prakoeswa, Cita Rosita S.; Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi
Indonesian Journal of Tropical and Infectious Disease Vol 6, No 1 (2016)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (785.454 KB) | DOI: 10.20473/ijtid.v6i1.1206

Abstract

Leprosy in twins is rarely reported. A 19 years-old male student, from Lamongan district, was diagnosed as Multibacillary (MB) leprosy in the Skin and STD Clinic of Dr. Soetomo General Hospital Surabaya. Multiple anesthetic skin lesions were found, but the bacteriologic examination was negative for Acid Fast Bacilli (AFB). Histopathology examination support the diagnosis of BL type of leprosy. His twin brother that has been lived together since born until present seems healthy without any complaints of skin lesions and have no signs of leprosy. When a serologic examination for leprosy was performed, a high anti PGL-1 antibody level was found in patient (IgM anti PGL-1 2937 and IgG anti PGL-1 3080 unit/ml) while his healthy twin brother showed only low level (IgM 745 and IgG 0 unit/ml). Interestingly when a PCR study was performed to detect M.leprae in the blood, both of them showed positive results. Using the TTC method, a genomic study of for M.leprae, it is revealed that both samples were identic ( 27x TTC repeats). According to patient’s history, he had a traffic accident and got a wound in the knee seven years ago, while the skin lesions seems started from this area around three years ago before it spread to other parts of the body. The patient was treated with Multi-drug therapy (MDT) while his sibling got a prophylactic treatment for leprosy. After 6 months of treatment, the leprosy skin lesions were diminished and the serologic anti PGL 1 has been decreased. His healthy brother also showed a decrease in anti PGL 1 level and no skin signs of leprosy.
COMPARATIVE STUDY ON THE INTENSITY OF Mycobacterium leprae EXPOSURE TO CHILDREN WHO LIVE IN LOW AND HIGH ALTITUDE IN LOW LEPROSY ENDEMIC AREA OF SOUTH SULAWESI Rachmawati, Rachmawati; Mataallo, Timurleng Tonang; Adam, Safruddin; Adam, A. M.; Amin, Safruddin; Tabri, Farida; Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi; Agusni, Indropo
Indonesian Journal of Tropical and Infectious Disease Vol 4, No 4 (2013)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1373.847 KB) | DOI: 10.20473/ijtid.v4i4.220

Abstract

Background: The intensity of Mycobacterium leprae exposure to people who live in leprosy endemic area could be measured by serological study and detection of the bacilli in the nose cavity. Different geographical altitude might have some influences to this exposure since the bacilli prefer to live in warm areas. Aim: A combined serological and PCR study of leprosy was conducted in Selayar island, South Sulawesi to 80 school children (40 from low land and 40 from highland altitudes) in order to compare the exposure intensity between the two areas. Method: Anti PGL-1 IgM antibody (ELISA) and PCR study to detect M.leprae in the nasal cavity were performed simultaneously from each person. Result: Seropositive cases were found in 23/40 children from low land compared to 16/40 children from high land, but statistically no significant difference (p>0.05). PCR positive for M.leprae in the nasal cavity only found in 1/40 children, both in low and high altitude. Conclusion: It is concluded that although the existence of M.leprae in nasal cavity is minimal, the intensity of exposure to this bacilli still high as indicated by serological study.
EVALUATION OF ANTI PCL-1 ANTIBODY TITER IN A GROUP OF HEALTHY SCHOOL CHILDREN WHO LIVE IN LEPROSY ENDEMIC AREA FROM 2007–2010 putri, rachmah diana; amiruddin, M. dali; tabri, farida; Adriaty, Dinar; Wahyuni, ratna; iswahyudi, iswahyudi; agusni, indropo; izumi, shinzo
Indonesian Journal of Tropical and Infectious Disease Vol 1, No 3 (2010)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (583.13 KB) | DOI: 10.20473/ijtid.v1i3.2184

Abstract

The “Iceberg phenomene” has been used to explain this situation which indicate that these new leprosy cases is originated from Subclinical Leprosy. Fifty eight healthy school children who live in Jeneponto Regency, a leprosy endemic area in South Sulawesi were recruited. The first examination was performed in 2007 and sera samples were kept in deep freeze refrigerator. In 2010 these children were re-examined for clinical leprosy and sera were collected again. ELISA study was performed simultaneously to these 58 pairs of sera (2007 & 2010) for measuring the titer of IgM anti PGL-1 antibody (ELISA) and the level 605u/ml was regarded as cut off value. After three years evaluation, none of these children showed any clinical signs of leprosy, but 20 of 22 ( 90.9%) children were remained sero-positive and only 2 (9.1%) became sero-negatives. In other sites, 5 children that previously sero-negatives became sero-positives after 3 years. Eight of 10 (80%) children who showed sero (+) with high titer (>1.000u/ml) in 2007, were also remained in high titer. The mean titer of 2007 was 627.8 u/ml, and after3 years became 723.9 u/ml (p<0.05). Although there is no progression from Subclinical to Manifest Leprosy cases among these children, the number of sero (+) cases were increased and the mean titer of IgM anti PGL-1 antibody was significantly increased.. The majority who previously showed high anti PGL-1 antibody titer, remained in high level. This study support the “iceberg phenomene” theory in Leprosy.
COMPARATIVE STUDY ON THE INTENSITY OF Mycobacterium leprae EXPOSURE BETWEEN HOUSEHOLD AND NONHOUSEHOLD CONTACT OF LEPROSY Arsyad, Yuniarti; Jifanti, Friska; Amiruddin, Muhammad Dali; Anwar, Anis Irawan; Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi; Agusni, Indropo
Indonesian Journal of Tropical and Infectious Disease Vol 3, No 1 (2012)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1081.524 KB) | DOI: 10.20473/ijtid.v3i1.192

Abstract

Leprosy stills a public health problem in West Sulawesi which has a Case Detection Rate (CDR) around 43.69/100.000 population. Household contacts of leprosy are a high risk group to be infected, due to droplet infection mode of transmission of the disease. A nose swab examination and serological study was conducted to detect exposure of M. leprae of people who live in leprosy endemic area. Detection of M. leprae in the nasal cavity will represent the exposure rate from outside and the measurement of specific antibody is represented the result of exposure to the immune system. Two group of inhabitants (30 household contacts of leprosy and 30 nonhousehold contacts) were involved in the study. They live in Banggae district, a leprosy endemic area of Majene Regency, West Sulawesi. Sixty nose swab samples and sixty capillary blood samples from the same invidividuals of the two groups were collected and sent to Leprosy laboratory of the Institute of Tropical Disease, Airlangga University Surabaya. A Polymerase Chain Reaction (PCR) was performed to the nose swab samples for detection of M. leprae. The blood samples were examined serologically to measure the level of anti PGL-1 antibody. PCR examination of nose swab samples showed 1/30 positive result in the household contact group and also 1/30 positive result in non-household contact of leprosy (statistically no significant difference, p > 0.05). Serological study showed higher sero-positive result in the household contact group (15/30 or 50%) compared to non-household contact (11/30 or 36%), but statistical calculation revealed no significant difference between the two groups (p > 0.05) on sero-positive results of leprosy. It is concluded that household and non-household contact in leprosy have the same risk to be affected by the disease. The term of household and non-household contact need to be redefined. The possible role of exposure from the environment was also discussed, especially from non-human resource of M. leprae.
TTC Repeats Variation of Mycobacterium leprae Isolates for Analysis of Leprosy Transmission in Leprosy Endemic Area in East Java, INDONESIA Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi; Agusni, Indropo; Izumi, Shinzo
Indonesian Journal of Tropical and Infectious Disease Vol 1, No 1 (2010)
Publisher : Institute of Topical Disease

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1494.939 KB) | DOI: 10.20473/ijtid.v1i1.3722

Abstract

East Java province still has some pocket of leprosy endemic areas. In order to solve the problem, molecular typing will make it feasible to study the transmission pattern of Mycobacterium leprae in leprosy endemic area. The present study is to analyze the presence of M.leprae DNA in the environment and to study variation number of TTC repeats and their distribution. Poteran Island is located in Madura, East Java and was chosen because this island has a high prevalence of leprosy and remains stable for the last five years. All samples were analyzed by PCR and the numbers of TTC repeats were confirmed by direct sequencing. Of all collected samples, 26.4% isolates of water resources (24); 61.9% nasal swabs (26); and 35.3% skin tissues (24) are positives. No statistically difference in the pattern distribution of TTC repeats between skin tissues of patients and nasal swab of households contact (p=0.594); also distribution of TTC repeats between skin tissues of leprosy patients and those of water resources (p=0.441); and distribution of TTC repeats between nasal swab of households contact with water resources (p=0.906). It means that the transmission of M.leprae in leprosy endemic area has closely related in 3 aspects: agent, host & environment.