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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2019.1
- 수록면
- 645 - 650 (6page)
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초록· 키워드
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Despite differences in virulence between strains of Toxoplasma gondii, rapid and accurate genotyping methods are lacking. In this study, a method was developed to detect and genotype T. gondii in food and environmental samples using PCR and a novel peptide nucleic acid (PNA) melting array. An alignment of genome sequences for T. gondii type I, II, and III obtained from NCBI was generated, and a single nucleotide polymorphism analysis was performed to identify targets for PCR amplification and a PNA melting array. Prior to the PNA melting array, conventional PCR was used to amplify GRA6 of T. gondii. After amplification, the PNA melting array was performed using two different PNA hybridization probes with fluorescent labels (FAM and HEX) and quenchers. Melting curves for each probe were used to determine genotypes and identify mutations. A 214-bp region of the GRA6 gene of T. gondii was successfully amplified by PCR. For all T. gondii strains (type I, II, and III) used to evaluate specificity, the correct genotypes were determined by the PNA melting array. Non-T. gondii strains, including 14 foodborne pathogens and 3 protozoan parasites, such as Giardia lamblia, Cryptosporidium parvum, and Entamoeba histolytica, showed no signal, suggesting that the assay has a high specificity. Although this is only a proof-of-concept study, the assay is promising for the fast and reliable genotyping of T. gondii from food and environmental samples.
목차
등록된 정보가 없습니다.
참고문헌 (20)
참고문헌 신청
Nielsen PE / 1994 / Peptide nucleic acid (PNA). A DNA mimic with a peptide backbone / Bioconjug. Chem 5:3~7
Egholm M / 1993 / PNA hybridizes to complementary oligonucleotides obeying the Watson-Crick hydrogen-bonding rules / Nature 365:566~568
Stender H / 2002 / PNA for rapid microbiology / J. Microbiol. Methods 48:1~17
Lehtola MJ / 2006 / Fluorescence in situ hybridization using peptide nucleic acid probes for rapid detection of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis in potable-water biofilms / Appl. Environ. Microbiol 72:848~853
Montoya JG / 2004 / Toxoplasmosis / Lancet (London, England) 363:1965~1976
Egholm M / 1993 / PNA hybridizes to complementary oligonucleotides obeying the Watson-Crick hydrogen-bonding rules / Nature 365:566~568
Stender H / 2002 / PNA for rapid microbiology / J. Microbiol. Methods 48:1~17
Lehtola MJ / 2006 / Fluorescence in situ hybridization using peptide nucleic acid probes for rapid detection of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis in potable-water biofilms / Appl. Environ. Microbiol 72:848~853
Montoya JG / 2004 / Toxoplasmosis / Lancet (London, England) 363:1965~1976
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