片形吸虫分子检测技术的研究进展
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摘要
片形吸虫病(Fascioliasis)是由片形属吸虫(Fasciola spp.)感染所引起的一种危害严重的人兽共患寄生虫病,病原主要包括大片吸虫(Fasciola gigantica)、肝片吸虫(Fasciola hepatica)及其中间型。该病在全球范围内流行,严重损害人及动物健康,威胁公共卫生安全,造成巨大的经济损失。尽管目前在片形吸虫病的免疫预防及新型药物制剂的研发方面已经取得一些进展,但是该病防控的前提是对片形吸虫病的准确诊断及检测。本文对近二十年来片形吸虫病的分子诊断与检测技术的研究进展进行总结,包括基于核糖体DNA、线粒体DNA及全基因组序列的片形吸虫分子鉴定,以及基于这些遗传标记进行PCR诊断、TaqMan探针实时定量PCR、环介导等温扩增试验(LAMP)等检测及诊断方法,以期为人和动物片形吸虫病的早期诊断及微量检测技术的研究提供科学参考。
Fascioliasis is a severe zoonotic parasitic disease caused by infection with Fasciola spp.,such as Fasciola gigantica and Fasciola hepatica and their intermediate form.Fasciola is one of the most important pathogens of domestic livestock and humans with a global distribution,causing significant socio-economic losses.Although some advances had been made in the development of immunization and new pharmaceutical preparations against fascioliasis,the core question of prevention and control was diagnosis and detection accurately.This paper summarized the research advance of molecular diagnostic and detection techniques for fascioliasis in the past two decades,including those identification techniques base on genetic markers in ribosomal DNA,mitochondrial DNA and whole genome sequences,PCR-based diagnostic techniques,such as the Taq Man probe-based real-time quantitative PCR,loop-mediated isothermal amplification assay(LAMP) were also presented,with the objective of providing reference for the development of early and rapid detection and diagnostic techniques for human and animal fascioliasis.
Fascioliasis is a severe zoonotic parasitic disease caused by infection with Fasciola spp.,such as Fasciola gigantica and Fasciola hepatica and their intermediate form.Fasciola is one of the most important pathogens of domestic livestock and humans with a global distribution,causing significant socio-economic losses.Although some advances had been made in the development of immunization and new pharmaceutical preparations against fascioliasis,the core question of prevention and control was diagnosis and detection accurately.This paper summarized the research advance of molecular diagnostic and detection techniques for fascioliasis in the past two decades,including those identification techniques base on genetic markers in ribosomal DNA,mitochondrial DNA and whole genome sequences,PCR-based diagnostic techniques,such as the Taq Man probe-based real-time quantitative PCR,loop-mediated isothermal amplification assay(LAMP) were also presented,with the objective of providing reference for the development of early and rapid detection and diagnostic techniques for human and animal fascioliasis.
引文
[1] CWIKLINSKI K,O'NEILL S M,DONNELLY S,et al.A prospective view of animal and human Fasciolosis[J].Parasite Immunology,2016,38(9):558-568.
[2] PENG M,ICHINOMIYA M,OHTORI M,et al.Molecular characterization of Fasciola hepatica,Fasciola gigantica,and aspermic Fasciola sp.in China based on nuclear and mitochondrial DNA[J].Parasitology Research,2009,105(3):809-815.
[3] ALVAREZ ROJAS C A,JEX A R,GASSER R B,et al.Techniques for the diagnosis of Fasciola infections in animals:room for improvement[J].Advances in Parasitology,2014,85:65-107.
[4] ZHANG X X,CONG W,ELSHEIKHA H M,et al.De novo transcriptome sequencing and analysis of the juvenile and adult stages of Fasciola gigantica[J].Infection,Genetics&Evolution,2017,51:33-40.
[5] CHEN J X,CHEN M X,AI L,et al.An outbreak of human Fascioliasis gigantica in Southwest China[J].Plos One,2013,8(8):e71520.
[6]刘倩,程娜,周岩,等.片形吸虫病研究进展[J].中国寄生虫学与寄生虫病杂志,2013,31(3):229-234.LIU Qian, CHENG Na, ZHOU Yan, et al.Research progress on fascioliasis[J].Chinese Journal of Parasitology and Parasitic Diseases,2013,31(3):229-234.(in Chinese)
[7]艾琳,陈木新,吕山,等.云南宾川地区牛羊片形吸虫感染调查及分子鉴定[J].热带医学杂志,2013,13(6):791-794.AI Lin,CHEN Mu-xin,LUShan,et al.Surveillance and molecular identification of Fasciola spp.from cattle and goats at Binchuan,Yunnan province[J].Journal of Tropical Medicine,2013,13(6):791-794.(in Chinese)
[8] HACARIZ O,AKGUN M,KAVAK P,et al.Comparative transcriptome profiling approach to glean virulence and immunomodulation-related genes of Fasciola hepatica[J].Bmc Genomics,2015,16(1):366.
[9] NOVOBILSKyA,HOGLUND J.First report of closantel treatment failure against Fasciola hepatica in cattle[J].International Journal for Parasitology Drugs&Drug Resistance,2015,5(3):172-177.
[10] LIU G H,GSAAER R B,YOUNG N D,et al.Complete mitochondrial genomes of the ‘intermediate form’ of Fasciola and Fasciola gigantica, and their comparison with F. hepatica[J].Parasites&Vectors,2014,7(1):150.
[11] OMAR M A,METWALLY A M,SULTAN K.Molecular and phylogenetic status of Fasciola sp.,of cattle in Qena,Upper Egypt[J].Pakistan Journal of Biological Sciences,2013,16(15):726-730.
[12] AI L,CHEN M X,ALASAAD S,et al.Genetic characterization,species differentiation and detection of Fasciola spp.by molecular approaches[J].Parasites&Vectors,2011,4(1):101.
[13] HUANG W Y,HE B,WANG C R,et al.Characterisation of Fasciola species from Mainland China by ITS-2 ribosomal DNA sequence[J].Veterinary Parasitology,2004,120(1):75-83.
[14] AGATSUMA T,ARAKAWA Y,IWAGAMI M,et al.Molecular evidence of natural hybridization between Fasciola hepatica,and F.gigantica[J].Parasitology International,2000,49(3):231-238.
[15]伍慧兰,庄远红,黄文.湖南省华支睾吸虫线粒体cox 1和nad 1基因的克隆及进化分析[J].中国兽医科学,2012,42(1):8-12.WU Hui-lan,ZHUANG Yuan-hong,HUANG Wen.Cloning and phylogenetic analysis of mitochondrial cox 1and nad 1 genes of Clonorchis sinensis isolates from Hunan Province[J].Chinese Veterinary Science,2012,42(1):8-12.(in Chinese)
[16]宋慧群,艾琳,莫伊梦,等.片形吸虫线粒体nad 1基因的序列分析及种系发育[J].中国兽医科学,2009,39(6):487-491.SONG Hui-qun,AI Lin,MO Yi-meng,et al.Sequencing of mitochondrial NADH dehydrogenase subunit 1(nad 1)gene among Fasciola spp.and phylogenetic reconstruction[J].Chinese Veterinary Science,2009,39(6):487-491.(in Chinese)
[17]郭田田.云南大理市片形吸虫的分子鉴定及部分线粒体DNA的序列研究[D].昆明:昆明医科大学,2016.GUO Tian-tian.The study of molecular identification and the partical of mitochondrial DNA sequence of Fasciola in Dali,Yunnan Province[D].Kunming:Kunming Medical University,2016.(in Chinese)
[18] AI L,DONG S J,ZHANG W Y,et al.Specific PCR-based assays for the identification of Fasciola species:their development,evaluation and potential usefulness in prevalence surveys[J].Annals of Tropical Medicine&Parasitology,2010,104(1):65-72.
[19] MAGALHAES K G,PASSOS L K,CARVALHO ODOS S.Detection of Lymnaea columella infection by Fasciola hepatica through multiplex-PCR[J].Memórias Do Instituto Oswaldo Cruz,2004,99(4):421-424.
[20] MAGALHAES K G,JANNOTTI-PASSOS L K,CALDEIRA R L,et al.Isolation and detection of Fasciola hepatica DNA in Lymnaea viatrix from formalin-fixed and paraffin-embedded tissues through multiplex-PCR[J].Veterinary Parasitology,2008,152(3-4):333-338.
[21] BUSTIN S A.Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays[J].Journal of Molecular Endocrinology,2000,25(2):169-193.
[22] PAPLI N,LANDT O,FLEISCHER C,et al.Evaluation of a TaqMan real-time PCR for the detection of Theileria parva in buffalo and cattle[J].Veterinary Parasitology,2011,175(3-4):356-359.
[23] ALASAAD S,SORIGUER R C,ABU-MADI M,et al.A TaqMan real-time PCR-based assay for the identification of Fasciola spp.[J].Veterinary Parasitology,2011,179(1-3):266-271.
[24] MARCILLA A,BARGUES M D,MAS-COMA S.A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica[J].Molecular&Cellular Probes,2002,16(5):327-333.
[25] ICHIKAWA M,ITAGAKI T.Discrimination of the ITS1types of Fasciola spp. based on a PCR-RFLP method[J].Parasitology Research,2010,106(3):757-761.
[26] ARYAEIPOUR M,ROUHANI S,BANDEHPOUR M,et al.Genotyping and phylogenetic analysis of Fasciola spp.isolated from sheep and cattle using PCRRFLP in Ardabil Province,Northwestern Iran[J].Iranian Journal of Public Health,2014,43(10):1364-1371.
[27] SARKARI B,PARHOODE M,ABDOLAHI KHABISI S,et al.Genetic diversity of Fasciola spp.isolates from northern part of Iran:comparison with southwestern isolates[J].Journal of Parasitic Diseases,2017,41(3):768-772.
[28] LIN R Q,DONG S J,NIE K,et al.Sequence analysis of the first internal transcribed spacer of rDNA supports the existence of the intermediate Fasciola between F.hepatica and F.gigantica in mainland China[J].Parasitology Research,2007,101(3):813-817.
[29] ALASAAD S,SORIGUER R C,ABU-MADI M,et al.A fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism(F-PCRSSCP)assay for the identification of Fasciola spp.[J].Parasitology Research,2011,108(6):1513-1517.
[30] SCARCELLA S,MIRANDA-MIRANDA E,SOLANA M V,et al.Approach to molecular characterization of different strains of Fasciola hepatica using random amplified polymorphic DNA polymerase chain reaction[J].Parasitology Research,2015,114(4):1341-1345.
[31] VILAS R,VAZQUEZ-PRIETO S,PANIAGUA E.Contrasting patterns of population genetic structure of Fasciola hepatica from cattle and sheep:Implications for the evolution of anthelmintic resistance[J].Infection Genetics&Evolution,2012,12(1):45-52.
[32] GUNASEKAR K R,TEWARI A K,SREEKUMAR C,et al.Elucidation of genetic variability among different isolates of Fasciola gigantica(giant liver fluke)using random-amplified polymorphic DNA polymerase chain reaction[J].Parasitology Research,2008,103(5):1075-1081.
[33] NOTOMI T,OKAYAMA H,MASUBUCHI H,et al.Loop-mediated isothermal amplification of DNA[J].Nucleic Acids Research,2000,28(12):E63.
[34] NAGAMINE K,HASE T,NOTOMI T.Accelerated reaction by loop-mediated isothermal amplification using loop primers[J].Molecular&Cellular Probes,2002,16(3):223-229.
[35] KUBOKI N, INOUE N, SAKURAI T, et al.Loop-mediated isothermal amplification for detection of African trypanosomes[J].Journal of Clinical Microbiology,2003,41(12):5517-5524.
[36] KARANIS P,THEKISOE O,KIOUPTSI K,et al.Development and preliminary evaluation of a Loop-Mediated isothermal amplification procedure for sensitive detection of Cryptosporidium oocysts in fecal and water samples[J].Applied&Environmental Microbiology,2007,73(17):5660-5662.
[37] XU J, RONG R, ZHANG H Q, et al.Sensitive and rapid detection of Schistosoma japonicum DNA by loopmediated isothermal amplification(LAMP)[J].International Journal for Parasitology,2010,40(3):327-331.
[38] AI L,LI C,ELSHEIKHA H M,et al.Rapid identification and differentiation of Fasciola hepatica and Fasciola gigantica by a loop-mediated isothermal amplification(LAMP)assay[J].Veterinary Parasitology,2010,174(3-4):228-233.
[39] MARTINEZ-VALLADARES M,ROJO-VAZQUEZ F A.Loop-mediated isothermal amplification(LAMP)assay for the diagnosis of fasciolosis in sheep and its application under field conditions[J].Parasites&Vectors,2016,9(1):73.
[40] ARIFIN M I,HOGLUND J,NOVOBILSKAA.Comparison of molecular and conventional methods for the diagnosis of Fasciola hepatica infection in the field[J].Veterinary Parasitology,2016,232:8-11.
[2] PENG M,ICHINOMIYA M,OHTORI M,et al.Molecular characterization of Fasciola hepatica,Fasciola gigantica,and aspermic Fasciola sp.in China based on nuclear and mitochondrial DNA[J].Parasitology Research,2009,105(3):809-815.
[3] ALVAREZ ROJAS C A,JEX A R,GASSER R B,et al.Techniques for the diagnosis of Fasciola infections in animals:room for improvement[J].Advances in Parasitology,2014,85:65-107.
[4] ZHANG X X,CONG W,ELSHEIKHA H M,et al.De novo transcriptome sequencing and analysis of the juvenile and adult stages of Fasciola gigantica[J].Infection,Genetics&Evolution,2017,51:33-40.
[5] CHEN J X,CHEN M X,AI L,et al.An outbreak of human Fascioliasis gigantica in Southwest China[J].Plos One,2013,8(8):e71520.
[6]刘倩,程娜,周岩,等.片形吸虫病研究进展[J].中国寄生虫学与寄生虫病杂志,2013,31(3):229-234.LIU Qian, CHENG Na, ZHOU Yan, et al.Research progress on fascioliasis[J].Chinese Journal of Parasitology and Parasitic Diseases,2013,31(3):229-234.(in Chinese)
[7]艾琳,陈木新,吕山,等.云南宾川地区牛羊片形吸虫感染调查及分子鉴定[J].热带医学杂志,2013,13(6):791-794.AI Lin,CHEN Mu-xin,LUShan,et al.Surveillance and molecular identification of Fasciola spp.from cattle and goats at Binchuan,Yunnan province[J].Journal of Tropical Medicine,2013,13(6):791-794.(in Chinese)
[8] HACARIZ O,AKGUN M,KAVAK P,et al.Comparative transcriptome profiling approach to glean virulence and immunomodulation-related genes of Fasciola hepatica[J].Bmc Genomics,2015,16(1):366.
[9] NOVOBILSKyA,HOGLUND J.First report of closantel treatment failure against Fasciola hepatica in cattle[J].International Journal for Parasitology Drugs&Drug Resistance,2015,5(3):172-177.
[10] LIU G H,GSAAER R B,YOUNG N D,et al.Complete mitochondrial genomes of the ‘intermediate form’ of Fasciola and Fasciola gigantica, and their comparison with F. hepatica[J].Parasites&Vectors,2014,7(1):150.
[11] OMAR M A,METWALLY A M,SULTAN K.Molecular and phylogenetic status of Fasciola sp.,of cattle in Qena,Upper Egypt[J].Pakistan Journal of Biological Sciences,2013,16(15):726-730.
[12] AI L,CHEN M X,ALASAAD S,et al.Genetic characterization,species differentiation and detection of Fasciola spp.by molecular approaches[J].Parasites&Vectors,2011,4(1):101.
[13] HUANG W Y,HE B,WANG C R,et al.Characterisation of Fasciola species from Mainland China by ITS-2 ribosomal DNA sequence[J].Veterinary Parasitology,2004,120(1):75-83.
[14] AGATSUMA T,ARAKAWA Y,IWAGAMI M,et al.Molecular evidence of natural hybridization between Fasciola hepatica,and F.gigantica[J].Parasitology International,2000,49(3):231-238.
[15]伍慧兰,庄远红,黄文.湖南省华支睾吸虫线粒体cox 1和nad 1基因的克隆及进化分析[J].中国兽医科学,2012,42(1):8-12.WU Hui-lan,ZHUANG Yuan-hong,HUANG Wen.Cloning and phylogenetic analysis of mitochondrial cox 1and nad 1 genes of Clonorchis sinensis isolates from Hunan Province[J].Chinese Veterinary Science,2012,42(1):8-12.(in Chinese)
[16]宋慧群,艾琳,莫伊梦,等.片形吸虫线粒体nad 1基因的序列分析及种系发育[J].中国兽医科学,2009,39(6):487-491.SONG Hui-qun,AI Lin,MO Yi-meng,et al.Sequencing of mitochondrial NADH dehydrogenase subunit 1(nad 1)gene among Fasciola spp.and phylogenetic reconstruction[J].Chinese Veterinary Science,2009,39(6):487-491.(in Chinese)
[17]郭田田.云南大理市片形吸虫的分子鉴定及部分线粒体DNA的序列研究[D].昆明:昆明医科大学,2016.GUO Tian-tian.The study of molecular identification and the partical of mitochondrial DNA sequence of Fasciola in Dali,Yunnan Province[D].Kunming:Kunming Medical University,2016.(in Chinese)
[18] AI L,DONG S J,ZHANG W Y,et al.Specific PCR-based assays for the identification of Fasciola species:their development,evaluation and potential usefulness in prevalence surveys[J].Annals of Tropical Medicine&Parasitology,2010,104(1):65-72.
[19] MAGALHAES K G,PASSOS L K,CARVALHO ODOS S.Detection of Lymnaea columella infection by Fasciola hepatica through multiplex-PCR[J].Memórias Do Instituto Oswaldo Cruz,2004,99(4):421-424.
[20] MAGALHAES K G,JANNOTTI-PASSOS L K,CALDEIRA R L,et al.Isolation and detection of Fasciola hepatica DNA in Lymnaea viatrix from formalin-fixed and paraffin-embedded tissues through multiplex-PCR[J].Veterinary Parasitology,2008,152(3-4):333-338.
[21] BUSTIN S A.Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays[J].Journal of Molecular Endocrinology,2000,25(2):169-193.
[22] PAPLI N,LANDT O,FLEISCHER C,et al.Evaluation of a TaqMan real-time PCR for the detection of Theileria parva in buffalo and cattle[J].Veterinary Parasitology,2011,175(3-4):356-359.
[23] ALASAAD S,SORIGUER R C,ABU-MADI M,et al.A TaqMan real-time PCR-based assay for the identification of Fasciola spp.[J].Veterinary Parasitology,2011,179(1-3):266-271.
[24] MARCILLA A,BARGUES M D,MAS-COMA S.A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica[J].Molecular&Cellular Probes,2002,16(5):327-333.
[25] ICHIKAWA M,ITAGAKI T.Discrimination of the ITS1types of Fasciola spp. based on a PCR-RFLP method[J].Parasitology Research,2010,106(3):757-761.
[26] ARYAEIPOUR M,ROUHANI S,BANDEHPOUR M,et al.Genotyping and phylogenetic analysis of Fasciola spp.isolated from sheep and cattle using PCRRFLP in Ardabil Province,Northwestern Iran[J].Iranian Journal of Public Health,2014,43(10):1364-1371.
[27] SARKARI B,PARHOODE M,ABDOLAHI KHABISI S,et al.Genetic diversity of Fasciola spp.isolates from northern part of Iran:comparison with southwestern isolates[J].Journal of Parasitic Diseases,2017,41(3):768-772.
[28] LIN R Q,DONG S J,NIE K,et al.Sequence analysis of the first internal transcribed spacer of rDNA supports the existence of the intermediate Fasciola between F.hepatica and F.gigantica in mainland China[J].Parasitology Research,2007,101(3):813-817.
[29] ALASAAD S,SORIGUER R C,ABU-MADI M,et al.A fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism(F-PCRSSCP)assay for the identification of Fasciola spp.[J].Parasitology Research,2011,108(6):1513-1517.
[30] SCARCELLA S,MIRANDA-MIRANDA E,SOLANA M V,et al.Approach to molecular characterization of different strains of Fasciola hepatica using random amplified polymorphic DNA polymerase chain reaction[J].Parasitology Research,2015,114(4):1341-1345.
[31] VILAS R,VAZQUEZ-PRIETO S,PANIAGUA E.Contrasting patterns of population genetic structure of Fasciola hepatica from cattle and sheep:Implications for the evolution of anthelmintic resistance[J].Infection Genetics&Evolution,2012,12(1):45-52.
[32] GUNASEKAR K R,TEWARI A K,SREEKUMAR C,et al.Elucidation of genetic variability among different isolates of Fasciola gigantica(giant liver fluke)using random-amplified polymorphic DNA polymerase chain reaction[J].Parasitology Research,2008,103(5):1075-1081.
[33] NOTOMI T,OKAYAMA H,MASUBUCHI H,et al.Loop-mediated isothermal amplification of DNA[J].Nucleic Acids Research,2000,28(12):E63.
[34] NAGAMINE K,HASE T,NOTOMI T.Accelerated reaction by loop-mediated isothermal amplification using loop primers[J].Molecular&Cellular Probes,2002,16(3):223-229.
[35] KUBOKI N, INOUE N, SAKURAI T, et al.Loop-mediated isothermal amplification for detection of African trypanosomes[J].Journal of Clinical Microbiology,2003,41(12):5517-5524.
[36] KARANIS P,THEKISOE O,KIOUPTSI K,et al.Development and preliminary evaluation of a Loop-Mediated isothermal amplification procedure for sensitive detection of Cryptosporidium oocysts in fecal and water samples[J].Applied&Environmental Microbiology,2007,73(17):5660-5662.
[37] XU J, RONG R, ZHANG H Q, et al.Sensitive and rapid detection of Schistosoma japonicum DNA by loopmediated isothermal amplification(LAMP)[J].International Journal for Parasitology,2010,40(3):327-331.
[38] AI L,LI C,ELSHEIKHA H M,et al.Rapid identification and differentiation of Fasciola hepatica and Fasciola gigantica by a loop-mediated isothermal amplification(LAMP)assay[J].Veterinary Parasitology,2010,174(3-4):228-233.
[39] MARTINEZ-VALLADARES M,ROJO-VAZQUEZ F A.Loop-mediated isothermal amplification(LAMP)assay for the diagnosis of fasciolosis in sheep and its application under field conditions[J].Parasites&Vectors,2016,9(1):73.
[40] ARIFIN M I,HOGLUND J,NOVOBILSKAA.Comparison of molecular and conventional methods for the diagnosis of Fasciola hepatica infection in the field[J].Veterinary Parasitology,2016,232:8-11.