Phần 1: Nghiên cứu sinh học phân tử trên các loại sán lá, đơn bào và mối liên quan đến ung thư cơ quan ở người

Trong bối cảnh biến đổi khí hậu toàn cầu (Global Climate change), nhiều bệnh truyền nhiễm đang nổi và tái nổi có xu hướng tăng lên đáng kể và đe dọa tử vong cũng như gia tăng gánh nặng bệnh tật toàn cầu. Trong đó một số bệnh ký sinh trùng và đơn bào có liên quan có ý nghĩa với một số bệnh ung thư như sán lá gan nhỏ Opisthorchis viverrini, Clonorchis sinensis liên quan đến ung thư biểu mô đường mật (cholangiocarcinoma), hay sán máng Schitosoma haematobium liên quan ung thư bàng quan tiết niệu, hay Cryptosporidium parvum có liên quan đến ung thư đại trực tràng (colorectal cancer).

Bên cạnh đó, việc xác định loài các giun sán về mặt hình thái học và sinh học phân tử phần nào làm rõ về phả hệ di truyền của các loài ký sinh trùng gây bệnh ở người. Các nghiên cứu sau đây làm rõ các vấn đề mà gần đây các nhà khoa học đang quan tâm làm rõ.

Molecular differentiation of Opisthorchis viverrini and Clonorchis sinensis eggs by multiplex real-time PCR with high resolution melting analysis.

Kaewkong W, Intapan PM, Sanpool O, Janwan P, Thanchomnang T, Laummaunwai P, Lulitanond V, Doanh PN, Maleewong W.

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Hình 1

Rapid detection and differentiation of Clonorchis sinensis and Opisthorchis viverrini eggs in human fecal samples using a duplex real-time fluorescence resonance energy transfer PCR and melting curve analysis.

Sanpool O, Intapan PM, Thanchomnang T, Janwan P, Lulitanond V, Doanh PN, Van Hien H, Dung do T, Maleewong W, Nawa Y.

Parasitol Res. 2012 Jul;111(1):89‐96. doi: 10.1007/s00436‐011‐2804‐7. Epub 2012 Jan 13.

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Cai XQ, Yu HQ, Li R, Yue QY, Liu GH, Bai JS, Deng Y, Qiu DY, Zhu XQ.

ScientificWorldJournal. 2014;2014:893981. doi: 10.1155/2014/893981. Epub 2014 Oct 19.

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Clonorchis sinensis and Opisthorchis viverrini: development of a mitochondrial-based multiplex PCR for their identification and discrimination.

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Exp Parasitol. 2006 Feb;112(2):109‐14. Epub 2005 Nov 28.

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Molecular identification of Clonorchis sinensis and discrimination with other opisthorchid liver fluke species using multiple Ligation-depended Probe Amplification (MLPA).

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Molecular discrimination of Opisthorchis-like eggs from residents in a rural community of central Thailand.

Buathong S, Leelayoova S, Mungthin M, Ruang‐Areerate T, Naaglor T, Suwannahitatorn P, Piyaraj P, Taamasri P, Tan‐Ariya P.

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Retrotransposon OV-RTE-1 from the carcinogenic liver fluke Opisthorchis viverrini: potential target for DNA-based diagnosis.

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Discrimination of O. viverrini, C. sinensis, H. pumilio and H. taichui using nuclear DNA-based PCR targeting ribosomal DNA ITS regions.

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A new PCR-based approach indicates the range of Clonorchis sinensis now extends to Central Thailand.

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[The reagents kit to detect Metyorchis biis, Opisthorchis viverrini and Clonorchis sinensis, Opisthorchis felineus–agents of opisthorchiasis using technique of polymerase chain reaction in real-time].

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Rapid detection of Opisthorchis viverrini and Strongyloides stercoralis in human fecal samples using a duplex real-time PCR and melting curve analysis.

Janwan P, Intapan PM, Thanchomnang T, Lulitanond V, Anamnart W, Maleewong W.

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A novel nuclear marker, Pm-int9, for phylogenetic studies of Opisthorchis felineus, Opisthorchis viverrini, and Clonorchis sinensis (Opisthorchiidae, Trematoda).

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Hình 4

Rapid detection of Opisthorchis viverrini copro-DNA using loop-mediated isothermal amplification (LAMP).

Arimatsu Y, Kaewkes S, Laha T, Hong SJ, Sripa B.

Parasitol Int. 2012 Mar;61(1):178‐82. doi: 10.1016/j.parint.2011.08.009. Epub 2011 Aug 16.

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Specific diagnosis of Opisthorchis viverrini using loop-mediated isothermal amplification (LAMP) targeting parasite microsatellites.

Arimatsu Y, Kaewkes S, Laha T, Sripa B.

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Co-infection with Opisthorchis viverrini and Haplorchis taichui detected by human fecal examination in Chomtong district, Chiang Mai Province, Thailand.

Wongsawad C, Phalee A, Noikong W, Chuboon S, Nithikathkul C.

Parasitol Int. 2012 Mar;61(1):56‐9. doi: 10.1016/j.parint.2011.10.003. Epub 2011 Oct 25.

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Clonorchis sinensis and Opisthorchis spp. in Vietnam: current status and prospects.

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Trans R Soc Trop Med Hyg. 2016 Jan;110(1):13‐20. doi: 10.1093/trstmh/trv103. Review.

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Opisthorchis viverrini and Haplorchis taichui: development of a multiplex PCR assay for their detection and differentiation using specific primers derived from HAT-RAPD.

Wongsawad C, Wongsawad P.

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Sequences and gene organization of the mitochondrial genomes of the liver flukes Opisthorchis viverrini and Clonorchis sinensis (Trematoda).

Cai XQ, Liu GH, Song HQ, Wu CY, Zou FC, Yan HK, Yuan ZG, Lin RQ, Zhu XQ.

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Development and evaluation of PCR methods based o­n cytochrome c oxidase subunit o­ne (cox1) and NADH dehydrogenase subunit o­ne gene (nad1) to detect Opisthorchis viverrini in human fecal samples.

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Parasitol Res. 2015 Sep;114(9):3547‐9. doi: 10.1007/s00436‐015‐4640‐7. Epub 2015 Aug 5.

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Development of a polymerase chain reaction applicable to rapid and sensitive detection of Clonorchis sinensis eggs in human stool samples.

Cho PY, Na BK, Choi KM, Kim JS, Cho SH, Lee WJ, Lim SB, Cha SH, Park YK, Pak JH, Lee HW, Hong SJ, Kim TS.

Pathog Glob Health. 2013 Jul;107(5):253‐9. doi: 10.1179/2047773213Y.0000000099.

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Development of a PCR assay and pyrosequencing for identification of important human fish-borne trematodes and its potential use for detection in fecal specimens.

Tantrawatpan C, Intapan PM, Thanchomnang T, Sanpool O, Janwan P, Lulitanond V, Sadaow L, Maleewong W.

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Hình 6

Unlocking the transcriptomes of two carcinogenic parasites, Clonorchis sinensis and Opisthorchis viverrini.

Young ND, Campbell BE, Hall RS, Jex AR, Cantacessi C, Laha T, Sohn WM, Sripa B, Loukas A, Brindley PJ, Gasser RB.

PLoS Negl Trop Dis. 2010 Jun 22;4(6):e719. doi: 10.1371/journal.pntd.0000719.

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Adult Opisthorchis felineus major protein fractions deduced from transcripts: comparison with liver flukes Opisthorchis viverrini and Clonorchis sinensis.

Pomaznoy M, Tatkov S, Katokhin A, Afonnikov D, Babenko V, Furman D, Brusentsov I, Belavin P, Najakshin A, Guselnikov S, Vasiliev G, Sivkov A, Prokhortchouk E, Skryabin K, Mordvinov V.

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Detection of Clonorchis sinensis in stool samples using real-time PCR.

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Development of mitochondrial loop-mediated isothermal amplification for detection of the small liver fluke Opisthorchis viverrini (Opisthorchiidae; Trematoda; Platyhelminthes).

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J Clin Microbiol. 2012 Apr;50(4):1178‐84. doi: 10.1128/JCM.06277‐11. Epub 2012 Feb 8.

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Improvement of PCR for detection of Opisthorchis viverrini DNA in human stool samples.

Duenngai K, Sithithaworn P, Rudrappa UK, Iddya K, Laha T, Stensvold CR, Strandgaard H, Johansen MV.

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PCR Diagnosis of Opisthorchis viverrini and Haplorchis taichui Infections in a Lao Community in an area of endemicity and comparison of diagnostic methods for parasitological field surveys.

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Opisthorchis viverrini: detection by polymerase chain reaction (PCR) in human stool samples.

Umesha KR, Kumar S, Parvathi A, Duenngai K, Sithithaworn P, Karunasagar I, Karunasagar I.

Exp Parasitol. 2008 Dec;120(4):353‐6. doi: 10.1016/j.exppara.2008.09.004. Epub 2008 Sep 9.

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The zoonotic, fish-borne liver flukes Clonorchis sinensis, Opisthorchis felineus and Opisthorchis viverrini.

Petney TN, Andrews RH, Saijuntha W, Wenz‐Mücke A, Sithithaworn P.

Int J Parasitol. 2013 Nov;43(12‐13):1031‐46. doi: 10.1016/j.ijpara.2013.07.007. Epub 2013 Aug 24. Review.

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Genetic comparison of liver flukes, Clonorchis sinensis and Opisthorchis viverrini, based o­n rDNA and mtDNA gene sequences.

Park GM.

Parasitol Res. 2007 Jan;100(2):351‐7. Epub 2006 Aug 11.

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Hình 8

Immunology and molecular biology of Opisthorchis viverrini infection.

Wongratanacheewin S, Sermswan RW, Sirisinha S.

Acta Trop. 2003 Nov;88(3):195‐207. Review.

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Discrimination of Opisthorchis viverrini from Haplorchis taichui using COI sequence marker.

Thaenkham U, Visetsuk K, Dung do T, Waikagul J.

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Rapid molecular detection of Opisthorchis viverrini in human fecal samples by real-time polymerase chain reaction.

Intapan PM, Thanchomnang T, Lulitanond V, Pongsaskulchoti P, Maleewong W.

Am J Trop Med Hyg. 2009 Nov;81(5):917‐20. doi: 10.4269/ajtmh.2009.09‐0275.

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Hình 9

Molecular evidence of Opisthorchis viverrini in infected bithyniid snails in the Lao People’s Democratic Republic by specific hybridization probe-based real-time fluorescence resonance energy transfer PCR method.

Sri‐Aroon P, Intapan PM, Lohachit C, Phongsasakulchoti P, Thanchomnang T, Lulitanond V, Hiscox A, Phompida S, Sananikhom P, Maleewong W, Brey PT.

Parasitol Res. 2011 Apr;108(4):973‐8. doi: 10.1007/s00436‐010‐2140‐3. Epub 2010 Nov 11.

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Opisthorchis viverrini: Gene expression profiling of carcinogenic adult liver fluke worms using 5′ SAGE.

Chutiwitoonchai N, Shen Y, Zheng H, Xiong H, Zhao G, Imtawil K, Intapan PM, Wongkham S, Wongkham C.

Exp Parasitol. 2008 Dec;120(4):306‐13. doi: 10.1016/j.exppara.2008.08.004. Epub 2008 Aug 24.

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First report and molecular identification of Opisthorchis viverrini infection in human communities from Lower Myanmar.

Aung WPP, Htoon TT, Tin HH, Thinn KK, Sanpool O, Jongthawin J, Sadaow L, Phosuk I, Rodpai R, Intapan PM, Maleewong W.

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Molecular differentiation of Schistosoma japonicum and Schistosoma mekongi by real‐time PCR with high resolution melting analysis.

Kongklieng A, Kaewkong W, Intapan PM, Sanpool O, Janwan P, Thanchomnang T, Lulitanond V, Sri‐Aroon P, Limpanont Y, Maleewong W.

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Hình 10

Early detection and estimation of infection burden by real‐time PCR in rats experimentally infected with Clonorchis sinensis.

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Copro‐DNA diagnosis of Opisthorchis viverrini and Haplorchis taichui infection in an endemic area of Lao PDR.

Sato M, Pongvongsa T, Sanguankiat S, Yoonuan T, Dekumyoy P, Kalambaheti T, Keomoungkhoun M, Phimmayoi I, Boupha B, Moji K, Waikagul J.

Southeast Asian J Trop Med Public Health. 2010 Jan;41(1):28‐35.

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A specific PCR assay for the diagnosis of Clonorchis sinensis infection in humans, cats and fishes.

Huang SY, Tang JD, Song HQ, Fu BQ, Xu MJ, Hu XC, Zhang H, Weng YB, Lin RQ, Zhu XQ.

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Development of a PCR‐based method for the detection of Opisthorchis viverrini in experimentally infected hamsters.

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Parasitology. 2001 Feb;122(Pt 2):175‐80.

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Hình 11

Assessment of the genetic distinctions of Opisthorchis felineus from O. viverrini and Clonorchis sinensis by ITS2 and CO1 sequences.

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[The differentiation of Opisthorchis eggs by the ultrastructure of their outer membranes].

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Application of environmental DNA analysis for the detection of Opisthorchis viverrini DNA in water samples.

Hashizume H, Sato M, Sato MO, Ikeda S, Yoonuan T, Sanguankiat S, Pongvongsa T, Moji K, Minamoto T.

Acta Trop. 2017 May;169:1‐7. doi: 10.1016/j.actatropica.2017.01.008. Epub 2017 Jan 17.

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Development of a real‐time PCR assay for the detection of Clonorchis sinensis DNA in gallbladder bile and stone samples from patients with cholecystolithiasis.

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Hình 12+13

Real‐time RT‐PCR high‐resolution melting curve analysis and multiplex RT‐PCR to detect and differentiate grapevine leafroll‐associated virus 3 variant groups I, II, III and VI.

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Real‐time fluorescence resonance energy transfer PCR with melting curve analysis for the detection of Opisthorchis viverrini in fish intermediate hosts.

Intapan PM, Thanchomnang T, Lulitanond V, Phongsaskulchoti P, Maleewong W.

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Genome size estimation of liver fluke Opisthorchis viverrini by real‐time polymerase chain reaction based method.

Kaewkong W, Imtawil K, Maleewong W, Intapan PM, Sri‐Aroon P, Wongkham S, Wongkham C.

Parasitol Int. 2012 Mar;61(1):77‐80. doi: 10.1016/j.parint.2011.06.025. Epub 2011 Jul 4.

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Field survey focused o­n Opisthorchis viverrini infection in five provinces of Cambodia.

Miyamoto K, Kirinoki M, Matsuda H, Hayashi N, Chigusa Y, Sinuon M, Chuor CM, Kitikoon V.

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Ultrastructural investigations o­n the effects of praziquantel o­n human trematodes from Asia: Clonorchis sinensis, Metagonimus yokogawai, Opisthorchis viverrini, Paragonimus westermani and Schistosoma japonicum.

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Microscopic examination of gallbladder stones improves rate of detection of Clonorchis sinensis infection.

Qiao T, Ma RH, Luo XB, Zheng PM, Luo ZL, Yang LQ.

J Clin Microbiol. 2013 Aug;51(8):2551‐5. doi: 10.1128/JCM.00946‐13. Epub 2013 May 22.

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Enzyme‐linked immunosorbent assay for detection of Opisthorchis viverrini infection.

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Identification and characterization of microRNAs in Clonorchis sinensis of human health significance.

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BMC Genomics. 2010 Sep 28;11:521. doi: 10.1186/1471‐2164‐11‐521.

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Hình 14

Cathepsin F cysteine protease of the human liver fluke, Opisthorchis viverrini.

Pinlaor P, Kaewpitoon N, Laha T, Sripa B, Kaewkes S, Morales ME, Mann VH, Parriott SK, Suttiprapa S, Robinson MW, To J, Dalton JP, Loukas A, Brindley PJ.

PLoS Negl Trop Dis. 2009;3(3):e398. doi: 10.1371/journal.pntd.0000398. Epub 2009 Mar 24.

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Genetic variation among Clonorchis sinensis isolates from different geographic regions in China revealed by sequence analyses of four mitochondrial genes.

Liu GH, Li B, Li JY, Song HQ, Lin RQ, Cai XQ, Zou FC, Yan HK, Yuan ZG, Zhou DH, Zhu XQ.

J Helminthol. 2012 Dec;86(4):479‐84. doi: 10.1017/S0022149X11000757. Epub 2011 Dec 12.

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Detection of Opisthorchis viverrini in infected bithynid snails by real‐time fluorescence resonance energy transfer PCR‐based method and melting curve analysis.

Intapan PM, Thanchomnang T, Lulitanond V, Pongsaskulchoti P, Maleewong W.

Parasitol Res. 2008 Aug;103(3):649‐55. doi: 10.1007/s00436‐008‐1026‐0. Epub 2008 May 30.

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Effect of artesunate and artemether against Clonorchis sinensis and Opisthorchis viverrini in rodent models.

Keiser J, Shu‐Hua X, Jian X, Zhen‐San C, Odermatt P, Tesana S, Tanner M, Utzinger J.

Int J Antimicrob Agents. 2006 Oct;28(4):370‐3. Epub 2006 Sep 14.

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Hình 15

Detection of Opisthorchis viverrini by monoclonal antibody‐based ELISA and DNA hybridization.

Sirisinha S, Chawengkirttikul R, Sermswan R, Amornpant S, Mongkolsuk S, Panyim S.

Am J Trop Med Hyg. 1991 Feb;44(2):140‐5.

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Application of a loop‐mediated isothermal amplification (LAMP) assay targeting cox1 gene for the detection of Clonorchis sinensis in human fecal samples.

Rahman SMM, Song HB, Jin Y, Oh JK, Lim MK, Hong ST, Choi MH.

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