• Title/Summary/Keyword: BIP(Back Inner Primer)

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Detection of Salmonella typhi by Loop-mediated Isothermal Amplification Assay

  • Jo, Yoon-Kyung;Lee, Chang-Yeoul
    • Biomedical Science Letters
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    • v.14 no.2
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    • pp.115-118
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    • 2008
  • Salmonella typhi is frequent causes of foodborne illness and its detection is important for monitoring disease progression. In this study, by using general PCR and novel LAMP (Loop Mediated Isothermal Amplification) assay, we evaluated the usefulness of LAMP assay for detection of Salmonella typhi. In this LAMP assay, forward inner primer (FIP) and back inner primer (BIP) was specially designed for recognizing target invA gene. Target DNA was amplified and visualized as ladder-like pattern of bands on agarose gel within 60 min under isothermal conditions at $65^{\circ}C$. When the sensitivity and reproducibility of LAMP were compared to general PCR, there was no difference of reproducibility but sensitivity of LAMP assay was more efficient than PCR (the detection limit of LAMP assay was 30 fg, while the PCR assay was 3 pg). These results indicate that the LAMP assay is a potential and valuable means for detection of Salmonella typhi, especially for its rapidity, simplicity and low cost.

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Detection of Mycobacterium Tuberculosis by Loop-Mediated Isothermal Amplification Assay (등온 증폭법을 이용한 결핵균의 빠른 검출 시스템 개발)

  • Ahn, Young-Chang;Nam, Youn-Hyoung;Park, Su-Min;Cho, Min-Ho;Seo, Jae-Won;Yoon, Il-Kyu;Park, Yong-Hyun;Jang, Won-Cheoul
    • Journal of the Korean Chemical Society
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    • v.52 no.3
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    • pp.273-280
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    • 2008
  • Mycobacterium tuberculosis (MTB) remains a major worldwide public health problem. In recent years, the incidence of MTB has been rising. Rapid and reliable diagnosis of Mycobacterium tuberculosis is essential to initiate correct treatment, avoid severe complications, and prevent transmission. LAMP was used to develop a rapid and sensitive laboratory diagnostic system for the MTB. In this research, the loop-mediated isothermal amplification method (LAMP) that amplifies DNA with high specificity and rapidity at an isothermal condition was evaluated for rapid detection of MTB. Undiluted DNA (2.10 × 106 copy/mL), 10-1, 10-2, 10-3, 10-4, 10-5 and 10-6 (copy/mL) of MTB DNA were amplified by PCR and LAMP to determine the sensitivity of the assay. At results, the LAMP assay reported here has the advantages of rapid amplification, high sensitivity, and high specificity and will be useful for rapid and reliable clinical diagnosis of MTB in hospital clinical laboratory.