• Title/Summary/Keyword: RCR-RFLP

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PCR-RFLP for Rapid Subtyping of Plasmodium vivax Korean Isolates

  • Kang, Jung-Mi;Lee, Jinyoung;Kim, Tae Im;Koh, Eun-Ha;Kim, Tong-Soo;Sohn, Woon-Mok;Na, Byoung-Kuk
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.159-165
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    • 2017
  • Vivax malaria reemerged in Korea in 1993 and the outbreak has been continued with fluctuating numbers of annual indigenous cases. Understanding the nature of the genetic population of Plasmodium vivax circulating in Korea is beneficial for the knowledge of the nationwide parasite heterogeneity and in the implementation of malaria control programs in the country. Previously, we analyzed polymorphic nature of merozoite surface protein-1 (MSP-1) and MSP-$3{\alpha}$ in Korean P. vivax population and identified the Korean P. vivax population has been diversifying rapidly, with the appearance of parasites with new genetic subtypes, despite the recent reduction of the disease incidence. In the present study, we developed simple PCR-RFLP methods for rapid subtyping of MSP-1 and MSP-$3{\alpha}$ of Korean P. vivax isolates. These PCR-RFLP methods were able to easily distinguish each subtype of Korean P. vivax MSP-1 and MSP-$3{\alpha}$ with high accuracy. The PCR-RFLP subtyping methods developed here would be easily applied to massive epidemiological studies for molecular surveillance to understand genetic population of P. vivax and to supervise the genetic variation of the parasite circulating in Korea.

PCR-RFLP patterns of three kinds of Metagonimus in Korea (국내에 존재하는 세 종류 메타고니무스속 흡충의 RCR-RFLP반응양상)

  • 유재란;정진성
    • Parasites, Hosts and Diseases
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    • v.35 no.4
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    • pp.271-276
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    • 1997
  • We tried to compare the three kinds oi Metagonimur species. M. Wokognulci, Ifetafonimus Miyata type, and M. tnknhashii, which were Know to be distributed in Korea with polymerase chain reaction based-restriction fragment length polymorphism (PCR- RFLP) patterns. We amplified the internal transcribed spacer 1 (ITSI) site of ribosomal RNA and mitochondrial cytochrome c oxidase I (mCOI) gene. The restriction patterns of ITSI gene loth Rsc I, ALu I and Msp I showed multiple fragmented bands of different sizes between three kinds of Metcgonimus. In case of mc01 gene, Rsc I and Alu I enzymes produced differentially fragmented band patterns. According to the parsimony analysis of PCR-RFLP patterns, the estimated genetic divergence between M Wokognwai and Metasoninus Miyata type was 0.034880, between Metusoninus Miyata type and M. tckc- hushii was 0.028098, between M. wokogawai and M. tnkahashii was 0.018179. It is suggested that Metasonimus Miyata type may be separate species and evolutionize at the older time than the other two species.

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Application of 16S rDNA PCR-RFLP Analysis for the Rapid Identification of Weissella Species (Weissella 속 유산균의 빠른 동정을 위한 16S rDNA PCR-RFLP 분석법의 적용)

  • Lee, Myeong-Jae;Cho, Kyeung-Hee;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.455-460
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    • 2010
  • A polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis was applied to detect and identify ten Weissella spp. frequently found in kimchi. The previously reported genus-specific primers designed from 16S rDNA sequences of Weissella spp. were adopted but PCR was performed at the increased annealing temperature by $4^{\circ}C$. The sizes of amplified PCR products and restricted fragments produced by AluI, MseI, and BceAI endonucleases were well correspond with the expected sizes. W. kandleri, W. koreensis, W. confusa, W. minor, W. viridescens, W. cibaria, and W. soli were distinguished by AluI and MseI and W. hellenica and W. paramesenteroides were identified by BceAI. W. thailandensis was distinguished when restriction pattern of other species was compared but identified by the single use of MspI.