DEVELOPME NT AND VALIDATION OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) FOR DETECTION OF HUMAN HCV GENOME
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Keywords
Isothermal, Amplifications, LAMP, Diagnosing, Complications
Abstract
Hepatitis C virus is considered a major reason of liver damaging, within estimated 180 million effected peoples worldwide. This study was conducted to diagnose the RNA of hepatitis C virus through isothermal amplification method (LAMP). Total genome of HCV different genotypes was aligned by MEGA CLUSTAL-W and different primers were designed by explorer version 4.0 software. All the extracted RNA of hepatitis C virus was collected from different laboratories located at different areas of Lahore, Pakistan. Extracted DNA of hepatitis B virus and other species was also included in this study to check the specificity of this technique. Persistence of hepatitis C virus was diagnosed by and isothermal amplification technique (LAMP) on dry heat block, incubator and thermo-cycler respectively. Ladder like pattern were obtained during this isothermal amplification for positive extracted RNA and no pattern was obtained for both positive and negative control samples in the population of Lahore, Pakistan. Significant difference was also observed in time consuming between LAMP and general PCR diagnosing technique. These results suggested that special measures should be adopted to diagnose the RNA of hepatitis C virus to reduce the incidence of complications of hepatitis C which increase the mortality among hepatitis C patients.
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H.M Ahsan Sardar
Department of Wildlife and Ecology, University of Veterinary and Animal Sciences Lahore, Pakistan.
Khalid Mahmood Anjum
Department of Wildlife and Ecology, University of Veterinary and Animal Sciences Lahore, Pakistan
Muhammad Imran
Department of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences Lahore, Pakistan.
Usman Ahmad
Department of Biochemistry and Biotechnology, University of the Punjab Lahore, Pakistan.
Mohsin Munawar
Department of Zoology, Government College University Lahore, Pakistan.
Ahmed Imtanan Khalid
Department of Chemistry, Science Research Complex Allama Iqbal Open University Islamabad, Pakistan
Farah Ejaz
Department of Animal Sciences, Quaid-e-Azam University Islamabad, Pakistan.
Majid khan
Department of Zoology, Government Postgraduate College, Timergara, Pakistan
Hamid ullah
Department of Zoology, Bahawalnagar Campus IUB, Bahawalpur, Pakistan