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Loop-Mediated Isothermal Amplification (LAMP) for the Rapid Diagnosis of Herpes Simplex Virus Type 1 (HSV-1)
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چکیده: (5548 مشاهده) |
| Background and Aims: considering difficulties in usual laboratory methods in detection of viral infections, improved DNA-based diagnostic techniques are more reliable. Loop mediated isothermal amplification method (LAMP) is a nucleic acid amplification method that amplifies DNA using six primers which has been developed to diagnose viruses as a rapid and high efficiency test. In this study, the LAMP was used for detection of Herpes simplex virus. Methods: The set of primers were designed, for detection of HSV-based on gG fragment. The genome of the virus was extracted from the culture supernatant of infected cells. LAMP technique was optimized in term of temperature, time and the ingredients used for test. For detection of HSV-1 infection, sensitivity and specificity of this technique were determined by various dilutions of virus and infected samples with HSV-2 that was taken from Day Hospital of Tehran. Results: The sensitivity and specificity of HSV-1 specific LAMP method, reached about 500 copies/tube and 99.9% respectively. Furthermore, both the agarose gel electrophoresis containing Ethidium Bromide (EtBr) and the turbidity assay directly detected HSV-1 virus LAMP products in reactions. Conclusion: In this study, the reliability of LAMP for detection of HSV- was approved therefore this rapid, accurate, and cost-effective detection and quantification method may perhaps be an investigative tool which can be valid for detection of target viral genome in clinical specimens in the absence of the necessary facilities for performing PCR. |
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متن کامل [PDF 227 kb]
(1379 دریافت)
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نوع مطالعه: پژوهشي |
موضوع مقاله:
عمومى
دریافت: 1393/8/16 | پذیرش: 1393/8/16 | انتشار: 1393/8/16
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