次世代シーケンシング解析を併用したNested Multiplex PCR法による、らい菌の薬剤耐性および型別の新規同時同定法
「はじめに」らい菌(Mycobacterium leprae)は抗酸菌の一種で, ハンセン病の主な原因菌である. WHOの報告によると, 2021年のハンセン病新規患者登録数は約14万人と報告されたが, ハンセン病新規患者数は依然として多い. ハンセン病の病型には少菌型と多菌型があるが, 特に少菌型ハンセン病患者からのスメア検査ではらい菌はほとんど検出されず, DNA診断が困難である. 薬剤耐性診断は, ダプソン耐性を決定する遺伝子(folP1), リファンピシン耐性を決定する遺伝子(rpoB)やキノロン耐性を決定する遺伝子(gyrA)の薬剤耐性決定領域(Drug resistance det...
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| Published in | 日本ハンセン病学会雑誌 Vol. 92; no. 2; pp. 47 - 52 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | Japanese |
| Published |
日本ハンセン病学会
2023
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1342-3681 1884-314X |
| DOI | 10.5025/hansen.92.47 |
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| Abstract | 「はじめに」らい菌(Mycobacterium leprae)は抗酸菌の一種で, ハンセン病の主な原因菌である. WHOの報告によると, 2021年のハンセン病新規患者登録数は約14万人と報告されたが, ハンセン病新規患者数は依然として多い. ハンセン病の病型には少菌型と多菌型があるが, 特に少菌型ハンセン病患者からのスメア検査ではらい菌はほとんど検出されず, DNA診断が困難である. 薬剤耐性診断は, ダプソン耐性を決定する遺伝子(folP1), リファンピシン耐性を決定する遺伝子(rpoB)やキノロン耐性を決定する遺伝子(gyrA)の薬剤耐性決定領域(Drug resistance determining region, DRDR)における塩基配列の変異有無によって判定される. らい菌の遺伝子型別では, らい菌ゲノム上の一塩基多型(single-nucleotide polymorphism, SNP)を利用して, 4つの主要なSNPタイプと16のサブタイプに分類され, らい菌のゲノム解析結果から, SNP型別と分離地域の相関関係が得られることが報告されている. |
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| AbstractList | 「はじめに」らい菌(Mycobacterium leprae)は抗酸菌の一種で, ハンセン病の主な原因菌である. WHOの報告によると, 2021年のハンセン病新規患者登録数は約14万人と報告されたが, ハンセン病新規患者数は依然として多い. ハンセン病の病型には少菌型と多菌型があるが, 特に少菌型ハンセン病患者からのスメア検査ではらい菌はほとんど検出されず, DNA診断が困難である. 薬剤耐性診断は, ダプソン耐性を決定する遺伝子(folP1), リファンピシン耐性を決定する遺伝子(rpoB)やキノロン耐性を決定する遺伝子(gyrA)の薬剤耐性決定領域(Drug resistance determining region, DRDR)における塩基配列の変異有無によって判定される. らい菌の遺伝子型別では, らい菌ゲノム上の一塩基多型(single-nucleotide polymorphism, SNP)を利用して, 4つの主要なSNPタイプと16のサブタイプに分類され, らい菌のゲノム解析結果から, SNP型別と分離地域の相関関係が得られることが報告されている. |
| Author | 森, 修一 岩尾, 泰久 阿戸, 学 中田, 登 |
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| References | 11） WHO. Guidelines for global surveillance of drug resistance in leprosy. World Health Organization, Geneva, Switzerland, 2009. 6） Cambau E, Perani E, Guillemin I, Jamet P, Ji B. Multidrug-resistance to dapsone, rifampicin, and ofloxacin in Mycobacterium leprae. Lancet 349: 103–104, 1997. 19） Truman RW, Singh P, Sharma R, Busso P, Rougemont J, Paniz-Mondolfi A, Kapopoulou A, Brisse S, Scollard DM, Gillis TP, Cole ST. Probable zoonotic leprosy in the southern United States. N Engl J Med 364: 1626-1633, 2011. 8） Shepard CC. A kinetic method for the study of activity of drugs against Mycobacterium leprae in mice. Int J Lepr Other Mycobact Dis 35: 429-435, 1967. 10） Tomioka H, Saito H. Therapeutic efficacy of some new quinolones and a combination of ofloxacin with rifampin against Mycobacterium leprae infection induced in athymic nude mice. Int J Lepr Other Mycobact Dis 61: 250-254, 1993. 4） Pettit JH, Rees RJ. Sulphone resistance in leprosy. An experimental and clinical study. Lancet 2: 673-674, 1964. 2） Monot M, Honoré N, Garnier T, Zidane N, Sherafi D, Paniz-Mondolfi A, Matsuoka M, Taylor GM, Donoghue HD, Bouwman A, Mays S, Watson C, Lockwood D, Khamesipour A, Dowlati Y, Jianping S, Rea TH, Vera-Cabrera L, Stefani MM, Banu S, Macdonald M, Sapkota BR, Spencer JS, Thomas J, Harshman K, Singh P, Busso P, Gattiker A, Rougemont J, Brennan PJ, Cole ST. Comparative genomic and phylogeographic analysis of Mycobacterium leprae. Nat Genet 41: 1282-1289, 2009. 16） Groathouse NA, Rivoire B, Kim H, Lee H, Cho SN, Brennan PJ, Vissa VD. Multiple polymorphic loci for molecular typing of strains of Mycobacterium leprae. J Clin Microbiol 42: 1666-1672, 2004. 18） Sharma R, Singh P, Loughry WJ, Lockhart JM, Inman WB, Duthie MS, Pena MT, Marcos LA, Scollard DM, Cole ST, Truman RW. Zoonotic leprosy in the Southeastern United States. Emerg Infect Dis 21: 2127-2134, 2015. 25） Marijke Braet S, Jouet A, Aubry A, Van Dyck-Lippens M, Lenoir E, Assoumani Y, Baco A, Mzembaba A, Cambau E, Vasconcellos SEG, Rigouts L, Suffys PN, Hasker E, Supply P, de Jong BC. Investigating drug resistance of Mycobacterium leprae in the Comoros: an observational deep-sequencing study. Lancet Microbe 3: e693-e700, 2022. 5） Jacobson RR, Hastings RC. Rifampin-resistant leprosy. Lancet 2: 1304-1305, 1976. 14） Matsuoka M, Maeda S, Kai M, Nakata N, Chae GT, Gillis TP, Kobayashi K, Izumi S, Kashiwabara Y. Mycobacterium leprae typing by genomic diversity and global distribution of genotypes. Int J Lepr Other Mycobact Dis 68: 121-128, 2000. 24） Yu G, Fadrosh D, Goedert JJ, Ravel J, Goldstein AM. 2015. Nested PCR biases in interpreting microbial community structure in 16S rRNA gene sequence datasets. PLoS One 10: e0132253. 21） Benjak A, Avanzi C, Singh P, Loiseau C, Girma S, Busso P, Fontes ANB, Miyamoto Y, Namisato M, Bobosha K, Salgado CG, da Silva MB, Bouth RC, Frade MAC, Filho FB, Barreto JG, Nery JAC, Bührer-Sékula S, Lupien A, Al-Samie AR, Al-Qubati Y, Alkubati AS, Bretzel G, Vera-Cabrera L, Sakho F, Johnson CR, Kodio M, Fomba A, Sow SO, Gado M, Konaté O, Stefani MMA, Penna GO, Suffys PN, Sarno EN, Moraes MO, Rosa PS, Baptista IMFD, Spencer JS, Aseffa A, Matsuoka M, Kai M, Cole ST. Phylogenomics and antimicrobial resistance of the leprosy bacillus Mycobacterium leprae. Nat Commun 9: 352, 2018. 23） Chen X, He J, Liu J, You Y, Yuan L, Wen Y. Nested PCR and the TaqMan SNP genotyping assay enhanced the sensitivity of drug resistance testing of Mycobacterium leprae using clinical specimens of leprosy patients. PLoS Negl Trop Dis 13: e0007946, 2019. 22） Truman RW, Singh P, Sharma R, Busso P, Rougemont J, Paniz-Mondolfi A, Kapopoulou A, Brisse S, Scollard DM, Gillis TP, Cole ST. Probable zoonotic leprosy in the southern United States. N Engl J Med 364: 1626–1633, 2011. 3） Iwao Y, Mori S, Ato M, Nakata N. Simultaneous Determination of Mycobacterium leprae Drug Resistance and Single-Nucleotide Polymorphism Genotype by Use of Nested Multiplex PCR with Amplicon Sequencing. J Clin Microbiol 59: e00814-21, 2021 7） Mori S, Yotsu RR, Suzuki K, Makino M, Ishii N. Present situation of leprosy in Japan, 2006-2010: analysis of drug resistance in new registered and relapsed cases by molecular biological methods. J Dermatol Sci 67: 192-4, 2012. 15） Shin YC, Lee H, Lee H, Walsh GP, Kim JD, Cho SN. Variable numbers of TTC repeats in Mycobacterium leprae DNA from leprosy patients and use in strain differentiation. J Clin Microbiol 38: 4535-4538, 2000. 17） Monot M, Honoré N, Garnier T, Araoz R, Coppée JY, Lacroix C, Sow S, Spencer JS, Truman RW, Williams DL, Gelber R, Virmond M, Flageul B, Cho SN, Ji B, Paniz-Mondolfi A, Convit J, Young S, Fine PE, Rasolofo V, Brennan PJ, Cole ST. On the origin of leprosy. Science 308: 1040-1042, 2005. 20） Singh P, Cole ST. Mycobacterium leprae: genes, pseudogenes and genetic diversity. Future Microbiol 6: 57-71, 2011. 12） Maeda S, Matsuoka M, Nakata N, Kai M, Maeda Y, Hashimoto K, Kimura H, Kobayashi K, Kashiwabara Y. Multidrug resistant Mycobacterium leprae from patients with leprosy. Antimicrob Agents Chemother 45: 3635-3639, 2001. 13） Williams DL, Gillis TP, Portaels F. Geographically distinct isolates of Mycobacterium leprae exhibit no genotypic diversity by restriction fragment-length polymorphism analysis. Mol Microbiol 4: 1653-1659, 1990. 9） Kohsaka K, Mori T, Ito T. Lepromatoid lesion developed in nude mouse inoculated with Mycobacterium leprae--animal transmission of leprosy. Repura 45: 177-87, 1976. 1） https://www.who.int/data/gho/data/themes/topics/leprosy-hansens-disease |
| References_xml | – reference: 17） Monot M, Honoré N, Garnier T, Araoz R, Coppée JY, Lacroix C, Sow S, Spencer JS, Truman RW, Williams DL, Gelber R, Virmond M, Flageul B, Cho SN, Ji B, Paniz-Mondolfi A, Convit J, Young S, Fine PE, Rasolofo V, Brennan PJ, Cole ST. On the origin of leprosy. Science 308: 1040-1042, 2005. – reference: 9） Kohsaka K, Mori T, Ito T. Lepromatoid lesion developed in nude mouse inoculated with Mycobacterium leprae--animal transmission of leprosy. Repura 45: 177-87, 1976. – reference: 16） Groathouse NA, Rivoire B, Kim H, Lee H, Cho SN, Brennan PJ, Vissa VD. Multiple polymorphic loci for molecular typing of strains of Mycobacterium leprae. J Clin Microbiol 42: 1666-1672, 2004. – reference: 21） Benjak A, Avanzi C, Singh P, Loiseau C, Girma S, Busso P, Fontes ANB, Miyamoto Y, Namisato M, Bobosha K, Salgado CG, da Silva MB, Bouth RC, Frade MAC, Filho FB, Barreto JG, Nery JAC, Bührer-Sékula S, Lupien A, Al-Samie AR, Al-Qubati Y, Alkubati AS, Bretzel G, Vera-Cabrera L, Sakho F, Johnson CR, Kodio M, Fomba A, Sow SO, Gado M, Konaté O, Stefani MMA, Penna GO, Suffys PN, Sarno EN, Moraes MO, Rosa PS, Baptista IMFD, Spencer JS, Aseffa A, Matsuoka M, Kai M, Cole ST. Phylogenomics and antimicrobial resistance of the leprosy bacillus Mycobacterium leprae. Nat Commun 9: 352, 2018. – reference: 2） Monot M, Honoré N, Garnier T, Zidane N, Sherafi D, Paniz-Mondolfi A, Matsuoka M, Taylor GM, Donoghue HD, Bouwman A, Mays S, Watson C, Lockwood D, Khamesipour A, Dowlati Y, Jianping S, Rea TH, Vera-Cabrera L, Stefani MM, Banu S, Macdonald M, Sapkota BR, Spencer JS, Thomas J, Harshman K, Singh P, Busso P, Gattiker A, Rougemont J, Brennan PJ, Cole ST. Comparative genomic and phylogeographic analysis of Mycobacterium leprae. Nat Genet 41: 1282-1289, 2009. – reference: 10） Tomioka H, Saito H. Therapeutic efficacy of some new quinolones and a combination of ofloxacin with rifampin against Mycobacterium leprae infection induced in athymic nude mice. Int J Lepr Other Mycobact Dis 61: 250-254, 1993. – reference: 25） Marijke Braet S, Jouet A, Aubry A, Van Dyck-Lippens M, Lenoir E, Assoumani Y, Baco A, Mzembaba A, Cambau E, Vasconcellos SEG, Rigouts L, Suffys PN, Hasker E, Supply P, de Jong BC. Investigating drug resistance of Mycobacterium leprae in the Comoros: an observational deep-sequencing study. Lancet Microbe 3: e693-e700, 2022. – reference: 5） Jacobson RR, Hastings RC. Rifampin-resistant leprosy. Lancet 2: 1304-1305, 1976. – reference: 7） Mori S, Yotsu RR, Suzuki K, Makino M, Ishii N. Present situation of leprosy in Japan, 2006-2010: analysis of drug resistance in new registered and relapsed cases by molecular biological methods. J Dermatol Sci 67: 192-4, 2012. – reference: 4） Pettit JH, Rees RJ. Sulphone resistance in leprosy. An experimental and clinical study. Lancet 2: 673-674, 1964. – reference: 11） WHO. Guidelines for global surveillance of drug resistance in leprosy. World Health Organization, Geneva, Switzerland, 2009. – reference: 24） Yu G, Fadrosh D, Goedert JJ, Ravel J, Goldstein AM. 2015. Nested PCR biases in interpreting microbial community structure in 16S rRNA gene sequence datasets. PLoS One 10: e0132253. – reference: 23） Chen X, He J, Liu J, You Y, Yuan L, Wen Y. Nested PCR and the TaqMan SNP genotyping assay enhanced the sensitivity of drug resistance testing of Mycobacterium leprae using clinical specimens of leprosy patients. PLoS Negl Trop Dis 13: e0007946, 2019. – reference: 20） Singh P, Cole ST. Mycobacterium leprae: genes, pseudogenes and genetic diversity. Future Microbiol 6: 57-71, 2011. – reference: 22） Truman RW, Singh P, Sharma R, Busso P, Rougemont J, Paniz-Mondolfi A, Kapopoulou A, Brisse S, Scollard DM, Gillis TP, Cole ST. Probable zoonotic leprosy in the southern United States. N Engl J Med 364: 1626–1633, 2011. – reference: 1） https://www.who.int/data/gho/data/themes/topics/leprosy-hansens-disease – reference: 8） Shepard CC. A kinetic method for the study of activity of drugs against Mycobacterium leprae in mice. Int J Lepr Other Mycobact Dis 35: 429-435, 1967. – reference: 15） Shin YC, Lee H, Lee H, Walsh GP, Kim JD, Cho SN. Variable numbers of TTC repeats in Mycobacterium leprae DNA from leprosy patients and use in strain differentiation. J Clin Microbiol 38: 4535-4538, 2000. – reference: 13） Williams DL, Gillis TP, Portaels F. Geographically distinct isolates of Mycobacterium leprae exhibit no genotypic diversity by restriction fragment-length polymorphism analysis. Mol Microbiol 4: 1653-1659, 1990. – reference: 14） Matsuoka M, Maeda S, Kai M, Nakata N, Chae GT, Gillis TP, Kobayashi K, Izumi S, Kashiwabara Y. Mycobacterium leprae typing by genomic diversity and global distribution of genotypes. Int J Lepr Other Mycobact Dis 68: 121-128, 2000. – reference: 3） Iwao Y, Mori S, Ato M, Nakata N. Simultaneous Determination of Mycobacterium leprae Drug Resistance and Single-Nucleotide Polymorphism Genotype by Use of Nested Multiplex PCR with Amplicon Sequencing. J Clin Microbiol 59: e00814-21, 2021 – reference: 19） Truman RW, Singh P, Sharma R, Busso P, Rougemont J, Paniz-Mondolfi A, Kapopoulou A, Brisse S, Scollard DM, Gillis TP, Cole ST. Probable zoonotic leprosy in the southern United States. N Engl J Med 364: 1626-1633, 2011. – reference: 6） Cambau E, Perani E, Guillemin I, Jamet P, Ji B. Multidrug-resistance to dapsone, rifampicin, and ofloxacin in Mycobacterium leprae. Lancet 349: 103–104, 1997. – reference: 12） Maeda S, Matsuoka M, Nakata N, Kai M, Maeda Y, Hashimoto K, Kimura H, Kobayashi K, Kashiwabara Y. Multidrug resistant Mycobacterium leprae from patients with leprosy. Antimicrob Agents Chemother 45: 3635-3639, 2001. – reference: 18） Sharma R, Singh P, Loughry WJ, Lockhart JM, Inman WB, Duthie MS, Pena MT, Marcos LA, Scollard DM, Cole ST, Truman RW. Zoonotic leprosy in the Southeastern United States. Emerg Infect Dis 21: 2127-2134, 2015. |
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| Snippet | 「はじめに」らい菌(Mycobacterium leprae)は抗酸菌の一種で, ハンセン病の主な原因菌である. WHOの報告によると, 2021年のハンセン病新規患者登録数は約14万人と報告されたが, ハンセン病新規患者数は依然として多い. ハンセン病の病型には少菌型と多菌型があるが,... |
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| SubjectTerms | nested multiplex PCR（nmPCR） Next-Generation Sequencing（NGS） single-nucleotide polymorphism（SNP） らい菌 |
| Title | 次世代シーケンシング解析を併用したNested Multiplex PCR法による、らい菌の薬剤耐性および型別の新規同時同定法 |
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| ispartofPNX | 日本ハンセン病学会雑誌, 2023, Vol.92(2), pp.47-52 |
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