DEVELOPME NT AND VALIDATION OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) FOR DETECTION OF HUMAN HCV GENOME

Main Article Content

H.M Ahsan Sardar
Khalid Mahmood Anjum
Muhammad Imran
Usman Ahmad
Mohsin Munawar
Ahmed Imtanan Khalid
Farah Ejaz
Majid khan
Hamid ullah

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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