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DAPSONE RESISTANCE IN A Mycobacterium leprae ISOLATE WITH TWO POINT MUTATIONS IN folP GENE FROM A LEPROSY PATIENT Ardiaty, Dinar; Wahyuni, Ratna; Prakoeswa, Cita; Abdullah, Rasyidin; Agusni, Indropo; Izumi, Shinzo
Indonesian Journal of Tropical and Infectious Disease Vol 3, No 2 (2012)
Publisher : Institute of Topical Disease

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Drug resistance in leprosy is important for Leprosy Control Program, since the WHO-Multidrug regiment (MDT) has been usedÂ for global treatment of leprosy for more than two decades already. A Dapsone resistance case in a Multibacillary (MB) leprosy caseÂ is reported. The patient was diagnosed and treated in Tajuddin Chalid Hospital Makassar, South Sulawesi. Previously he was treatedÂ in a health center at South Sulawesi and was given a treatment for one year, before referred to the hospital. The leprosy skin lesionsÂ are still active with erythematous skin lesions and thickened ear lobe. Bacteriological examination was positive for Acid Fast Bacilli,Â the Bacterial Index was 3+ and the Morphological Index was 1%. The specimens of M.leprae isolation was sent to the Institute ofÂ Tropical Disease Surabaya for drug resistance study. Using the Lp1-2 and Lp3-4 nested primers, PCR test was positive for M.leprae.Sequencing result for folP gene showed a double mutation at codon 53 (ACC / Threonin ) which become (AGG / Arginine). SimultaneousÂ mutation at two nucleotides at one codon has never been reported in Indonesia before and this phenomenon is important for leprosyÂ control policy.

Variation of TTC Repeat Pattern In The Dna of Mycobacterium Leprae Isolates Obtained from Archeological Bones and Leprosy Patients From East Nusa Tenggara Adriaty, Dinar; Wahyuni, Ratna; Iswahyudi, Iswahyudi; Aksono, Bimo; Koesbardiati, Toetik; Agusni, Indropo; Izumi, Shinzo
Journal of Tropical Life Science Vol 2, No 3 (2012)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.2.3.%x

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).

DAPSONE RESISTANCE IN A Mycobacterium leprae ISOLATE WITH TWO POINT MUTATIONS IN folP GENE FROM A LEPROSY PATIENT Ardiaty, Dinar; Wahyuni, Ratna; Prakoeswa, Cita; Abdullah, Rasyidin; Agusni, Indropo; Izumi, Shinzo
Indonesian Journal of Tropical and Infectious Disease Vol. 3 No. 2 (2012)
Publisher : Institute of Topical Disease Universitas Airlangga

Drug resistance in leprosy is important for Leprosy Control Program, since the WHO-Multidrug regiment (MDT) has been used for global treatment of leprosy for more than two decades already. A Dapsone resistance case in a Multibacillary (MB) leprosy case is reported. The patient was diagnosed and treated in Tajuddin Chalid Hospital Makassar, South Sulawesi. Previously he was treated in a health center at South Sulawesi and was given a treatment for one year, before referred to the hospital. The leprosy skin lesions are still active with erythematous skin lesions and thickened ear lobe. Bacteriological examination was positive for Acid Fast Bacilli, the Bacterial Index was 3+ and the Morphological Index was 1%. The specimens of M.leprae isolation was sent to the Institute of Tropical Disease Surabaya for drug resistance study. Using the Lp1-2 and Lp3-4 nested primers, PCR test was positive for M.leprae.Sequencing result for folP gene showed a double mutation at codon 53 (ACC / Threonin ) which become (AGG / Arginine). Simultaneous mutation at two nucleotides at one codon has never been reported in Indonesia before and this phenomenon is important for leprosy control policy.

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 Universitas Airlangga

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.

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 Universitas Airlangga

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.

Histoid Leprosy Rinasari, Umi; Sawitri, Sawitri; Listiawan, M. Yulianto; Prakoeswa, Cita Rosita; Agusni, Indropo; Santoso, Rachmat; Izumi, Shinzo
Indonesian Journal of Tropical and Infectious Disease Vol. 1 No. 1 (2010)
Publisher : Institute of Topical Disease Universitas Airlangga

Histoid Leprosy is a variant of lepromatous leprosy with characteristic clinical and histopathological features. Usually it is occured in lepromatous patients who relaps after dapsone monotherapy, in those with dapsone resistance , sometimes even after multidrug treatment, or at times, de novo with characteristic clinical and histopathological features. A 36 years old male, originated from Papua, visited to the skin outpatient clinic with translucent shiny nodules on the left elbow and thumb for the last 18 months. The nodules were multiple, painless and firm. There were nasal congestion, tickening of ear lobes and loss of eye brows. Patient did not have any history of previous antileprotic treatment. Routine blood examination was normal. Bacteriological examination of slit skin smear revealed acid-fast bacilli of Bacterial Index 4+ and Morfologic Index 10%. Histopathology of skin suggested lepromatous leprosy of histoid type with characteristic interlacing bundles of spindle shaped cells. Anti-PGL1 antibody (ELISA) revealed high titer of IgM (>5.300 u/ml) and also IgG anti PGL-1 (>5.300 u/ml). Polymerase chain reaction examination test to detect M.leprae was positive and direct sequencing of M.leprae isolate shows no mutation, which means no resistancy to MDT treatment. Treatment with MDTWHO regiment give clinical improvements and the histoid lesions disappered after 3 months treatment.The histoid form of leprosy in this case developed without any prior treatment of anti leprotic drugs ( de novo ). Some theoretical aspects of the patho-mechanism of histoid leprosy are discussed.

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 Universitas Airlangga

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.

Mycobacterium leprae Survival Inside Acanthamoeba sp. Isolated from Water Source in Leprosy Endemic Area, Indonesia: Mycobacterium leprae Survival Inside Acanthamoeba sp. Wahyuni, Ratna; Astari, Linda; Iswahyudi, Iswahyudi; Paling, Sepling; Adriaty, Dinar; Kurniawati, Siti; Aulia, Syifa; Rahmatari, Bandaru; Prakoeswa, Cita Rosita; Agusni, Indropo; Izumi, Shinzo
Journal of Tropical Life Science Vol. 14 No. 2 (2024)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.14.02.07

Mycobacterium leprae is an acid-fast bacterium that causes leprosy diseases, which remains a problem worldwide. Even though leprosy prevalence in the world has decreased significantly, many endemic pocket areas continue reporting new cases and harbor M. leprae in the environment, including water and soil. The presence of obligate intracellular bacteria-M. leprae in the environment raises a question on how it survives. Free-living amoeba has been proposed as its reservoir host in the environment. The study was conducted to give evidence that M. leprae can survive inside free-living amoeba isolated from water sources of leprosy endemic areas. M. leprae from leprosy patients was cultured together with Acanthamoeba sp. isolated from the water source of the leprosy endemic area. Viability and duplication of M. leprae inside amoeba then observed at day 14 and 28 using reverse transcriptase PCR and qPCR. The results showed that M. leprae survived inside the amoeba until day 28, but no bacterial replication was observed. The study reveals in vitro evidence of viable M. leprae inside free-living amoeba of leprosy endemic area environment.

Mycobacterium leprae Survival Inside Acanthamoeba sp. Isolated from Water Source in Leprosy Endemic Area, Indonesia: Mycobacterium leprae Survival Inside Acanthamoeba sp. Wahyuni, Ratna; Astari, Linda; Iswahyudi, Iswahyudi; Paling, Sepling; Adriaty, Dinar; Kurniawati, Siti; Aulia, Syifa; Rahmatari, Bandaru; Prakoeswa, Cita Rosita; Agusni, Indropo; Izumi, Shinzo
Journal of Tropical Life Science Vol. 14 No. 2 (2024)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.14.02.07

Mycobacterium leprae is an acid-fast bacterium that causes leprosy diseases, which remains a problem worldwide. Even though leprosy prevalence in the world has decreased significantly, many endemic pocket areas continue reporting new cases and harbor M. leprae in the environment, including water and soil. The presence of obligate intracellular bacteria-M. leprae in the environment raises a question on how it survives. Free-living amoeba has been proposed as its reservoir host in the environment. The study was conducted to give evidence that M. leprae can survive inside free-living amoeba isolated from water sources of leprosy endemic areas. M. leprae from leprosy patients was cultured together with Acanthamoeba sp. isolated from the water source of the leprosy endemic area. Viability and duplication of M. leprae inside amoeba then observed at day 14 and 28 using reverse transcriptase PCR and qPCR. The results showed that M. leprae survived inside the amoeba until day 28, but no bacterial replication was observed. The study reveals in vitro evidence of viable M. leprae inside free-living amoeba of leprosy endemic area environment.

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