scholarly journals Molecular detection of drug resistance in Mycobacterium leprae

2004 ◽  
Vol 75 (2) ◽  
pp. 118-130 ◽  
Author(s):  
Diana L. Williams ◽  
Thomas P. Gillis
Author(s):  
Yasuhisa Iwao ◽  
Shuichi Mori ◽  
Manabu Ato ◽  
Noboru Nakata

Mycobacterium leprae is the predominant cause of leprosy worldwide, and its genotypes can be classified into four single nucleotide polymorphism (SNP) types and 16 subtypes. Determining M. leprae drug resistance and genotype is typically done by PCR and Sanger DNA sequencing, which require substantial effort. Here we describe a rapid method involving multiplex PCR in combination with nested amplification and next generation sequence analysis that allows simultaneous determination of M. leprae drug resistance and SNP genotype directly from clinical specimens. We used this method to analyze clinical samples from two paucibacillary, nine multibacillary, and six type-undetermined leprosy patients. Regions in folP1 , rpoB , gyrA , and gyrB that determine drug resistance and those for 84 SNP-InDels in the M. leprae genome were amplified from clinical samples and their sequences were determined. The results showed that seven samples were subtype 1A, three were 1D, and seven were 3K. Three samples of the subtype 3K had folp1 mutation. The method may allow more rapid genetic analyses of M. leprae in clinical samples.


2005 ◽  
Vol 50 (1) ◽  
pp. 6-11 ◽  
Author(s):  
Eun-Young You ◽  
Tae Jin Kang ◽  
Se-Kon Kim ◽  
Seong-Beom Lee ◽  
Gue-Tae Chae

2011 ◽  
Vol 55 (6) ◽  
pp. 2971-2973 ◽  
Author(s):  
Pushpendra Singh ◽  
Philippe Busso ◽  
Alberto Paniz-Mondolfi ◽  
Nacarid Aranzazu ◽  
Marc Monot ◽  
...  

ABSTRACTPossible drug resistance inMycobacterium lepraestrains from Venezuela and three other South American countries was surveyed by molecular methods. None of the 230 strains from new leprosy cases exhibited drug resistance-associated mutations. However, two of the three strains from relapsed cases contained dapsone resistance mutations, and one strain also harbored a rifampin resistance mutation. Single nucleotide polymorphism analysis of these strains revealed five subtypes: 3I (73.8%), 4P (11.6%), 1D (6.9%), 4N (6%), and 4O (1.7%).


2009 ◽  
Vol 3 (3) ◽  
pp. 115-120 ◽  
Author(s):  
DINAR ADRIATY ◽  
RATNA WAHYUNI ◽  
CITA ROSITA S PRAKOESWA ◽  
NI PUTU SUSARI ◽  
INDROPO AGUSNI ◽  
...  

2018 ◽  
Vol 9 (1) ◽  
pp. 31-36
Author(s):  
Yustinus Maladan ◽  
C. S. Whinie Lestari ◽  
Ratna Tanjung ◽  
Vatim Dwi Cahyani ◽  
Muhammad Fajri Rokhmad

Abstrak Latar belakang: Lepra masih menjadi masalah kesehatan di Papua terutama di Kota Jayapura.Banyaknya kasus relaps dan default juga menjadi tantangan dalam eliminasi lepra di Jayapura. Kasusrelaps dan riwayat default pada beberapa penelitian berkaitan dengan resistensi terhadap multi drugtreatment (MDT). Tujuan penelitian ini adalah mendeteksi keberadaan mutasi gen rpoB M. leprae padapasien relaps, default dan pasein yang kurang peka terhadap terapi MDT di Kota Jayapura. Metode: Sampel diperoleh dari pasien yang terdiagnosis lepra dengan kriteria pasien relaps, default danpasien yang terus bergejala setelah terapi MDT sebanyak 34 sampel. Sampel diambil dalam bentuk insisikulit (skin silt) daun telinga. DNA diekstraksi dengan menggunakan kit Qiagen. Gen rpoB diamplifikasimelalui teknik PCR dan analisis nukleotida dilakukan melalui sekuensing. Analisis mutasi dilakukanmelalui BLAST dengan basis data GenBank. Hasil: Sebanyak 34 sampel yang diperiksa, 9 diantaranya positif BTA sedangkan 25 yang lainnya negatifBTA. Pada hasil PCR, sampel yang berhasil teramplifikasi sebanyak 31 sampel, dan 3 sampel tidakteramplifikasi. Hasil BLAST menunjukkan bahwa tidak ditemukan adanya mutasi pada gen rpoB yangdapat menyebabkan resistensi terhadap rifampisin. Kesimpulan: Kesimpulan dari penelitian ini adalah gen rpoB Mycobacterium leprae asal Jayapuratidak mengandung mutasi yang dapat menyebabkan terjadinya resistensi terhadap rifampisin. Dengandemikian rifampisin masih sensitif untuk pengobatan lepra di Kota Jayapura. Kata kunci: Lepra, gen rpoB, rifampisin, Mycobacterium leprae. AbstractBackground: Leprosy remains a prominent health problem in Papua especially in Jayapura City. Numerouscases of relapse and default are also challenges in leprosy elimination in Jayapura. Studies in Relapsecases and history of defaults revealed some resistance related to multi-drug treatment (MDT). The purposeof this study was to detect the presence of mutation in rpoB M. leprae gene in patient relapse, default andpatients who are less sensitive to MDT therapy in Jayapura City. Method: Samples were obtained from patients diagnosed with leprosy with criteria of relapse, defaultand symptomatic patients after receiving MDT therapy. A total of 34 samples were taken in the form ofskin incision (skin silt) of the earlobe. DNA was extracted using Qiagen kit. rpoB gene from extractedDNA was amplified through PCR method followed by nucleotide sequences. Analysis of mutation waselaborated using BLAST according to GenBank database. Result: 34 samples were examined, and 9 were positive for Ziehl-Neelsen (ZN)staining, while the 25 werenegative. In the PCR results, the samples that successfully amplified were 31 samples, and 3 samples werenot amplified. The results of BLAST indicated that no mutations in the rpoB gene found in which able toinitiate resistance to rifampicin. Conclusion: The conclusion of this study is the rpoB Mycobacterium leprae gene from Jayapura did notcontain any mutations that could trigger resistance to rifampicin. Thus rifampicin is still sensitive forleprosy treatment in Jayapura City. Keywords: Leprosy, rpoB gene, rifampicin, Mycobacterium leprae


2007 ◽  
Vol 78 (4) ◽  
pp. 343-352 ◽  
Author(s):  
Masanori Matsuoka ◽  
Teky Budiawan ◽  
Khin Saw Aye ◽  
Kyaw Kyaw ◽  
Esterlina Virtudes Tan ◽  
...  

2019 ◽  
Vol 70 (10) ◽  
pp. 2054-2061 ◽  
Author(s):  
Patrícia S Rosa ◽  
Helena R S D’Espindula ◽  
Ana C L Melo ◽  
Amanda N B Fontes ◽  
Amanda J Finardi ◽  
...  

AbstractBackgroundLeprosy has been treated with multidrug therapy, which has been distributed for free across the globe and regarded as highly efficient. However, the impossibility of growing Mycobacterium leprae in axenic media has historically impaired assessments of M. leprae resistance, a parameter only recently detectable through molecular methods.MethodsA systematic, population-based search for M. leprae resistance in suspected leprosy relapse cases and contacts was performed in Prata Village, an isolated, hyperendemic, former leprosy colony located in the Brazilian Amazon. Results led to an extended active search involving the entire Prata population. Confirmed leprosy cases were investigated for bacterial resistance using a combination of in vivo testing and direct sequencing of resistance genes folP1, rpoB, and gyrA. A molecular epidemiology analysis was performed using data from 17 variable number tandem repeats (VNTR).ResultsMycobacterium leprae was obtained from biopsies of 37 leprosy cases (18 relapses and 19 new cases): 16 (43.24%) displayed drug-resistance variants. Multidrug resistance to rifampicin and dapsone was observed in 8 relapses and 4 new cases. Single resistance to rifampicin was detected in 1 new case. Resistance to dapsone was present in 2 relapses and 1 new case. Combined molecular resistance and VNTR data revealed evidence of intra-familial primary transmission of resistant M. leprae.ConclusionsA comprehensive, population-based systematic approach to investigate M. leprae resistance in a unique population revealed an alarming scenario of the emergence and transmission of resistant strains. These findings may be used for the development of new strategies for surveillance of drug resistance in other populations.


2015 ◽  
Vol 21 (12) ◽  
pp. e85-e86 ◽  
Author(s):  
M. Lavania ◽  
A. Nigam ◽  
R.P. Turankar ◽  
I. Singh ◽  
P. Gupta ◽  
...  

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