• Title/Summary/Keyword: Nacleotide sequence homology

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Detection of Salmonella species by polymerase chain reaction (Polymerase chain reaction에 의한 Salmonella 속균의 검출)

  • Park, Doo-hee;Kim, Won-yong;Kim, Chul-joong;Mah, Jum-sool
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.115-125
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    • 1994
  • In this study, we try to establish the rapid and specific detection system for Salmonella species. The PhoE gene of Salmonella species was amplified with two specific primers, ST5 and ST8c, using PCR. The probe prepared from the amplified PhoE gene was sequenced and applied for Southern blot analysis. After PCR with ST5 and ST8c primers for PhoE gene, DNA bands of expected size(365bp) from 7 different Salmonella species were detected, but not from 12 enterobacteriaceae and 3 gram positive bacteria. PCR was highly sensitive to detect up to 10fg of purified DNA template and to identify Salmonella species with only 320 heat-lysed bacterial cells. The inhibition of PCR amplification from stool specimen was occurred with 50-fold dilution but disappeared over 100 fold dilution of samples. It was confirmed that the PhoE genes were amplified and cloned with over 97% nacleotide sequence homology of PCR products compared with that of S. typhfmurium LT2. The DNA probe derived from S. typhimurium TA 3,000 showed highly specific and sensitive reaction with PCR products of all tested Salmonella species. These results indicate that PCR was rapid and sensitive detection method for Salmonella species and DNA probe prepared from S. typhimurium TA 3,000 was specific to identify PCR products of different Salmonella species.

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Molecular Characterization of Hantavirus Isolates from Bandicota indica Captured in Indonesia and Thailand (인도네시아와 태국의 Bandicota indica 폐장조직에서 분리된 한타바이러스의 분자생물학적 특징)

  • Chu, Yong-Kyu;Cui, Longzhu;Song, Dae-Yong;Woo, Young-Dae;Praseno, Praseno;Leitmeyer, Katrin;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.30 no.3
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    • pp.203-210
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    • 2000
  • Hantaviruses are etiologic agents of hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS) in the world. Various hantaviruses were isolated from HFRS patients and several different rodent species in the world. Four hantavirus isolates from Indonesia and three isolates from Thailand among 89 Bandicotas captured in Yogyakarta, east region of Sumatra island, Indonesia and at Chiang Mai in Thailand during 1996 were made through several passages in Vero E6 cells. Viral genome M segment from two Indonesian isolates and three Thailand isolates were amplified using hantavirus generic primers of the M segment and cloned into pCRII vector. The genetic differences were analyzed by comparison of partial sequence of the M segment and antigenic differences were made by IFA. Nucleotide sequence homology of two isolates BC 8, BC 34 from Indonesia and two isolates thai 1322, thai 1330 to Seoul virus was 99% and 96%, respectively, but Thai 1164 was 80%Thai 1164 strain has shown 95% homology to Thai 749 virus. In conclusion it is indicated that two different serotype hantaviruses, Seoul and Thailand, are cocirculating among Bandicota in Thailand, in contrast Seoul serotype virus is circulating in Indonesia.

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