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Title

DEVELOPMENT AND VALIDATION OF A SIMPLE, RAPID AND INEXPENSIVE PCRRFLP METHOD FOR GENOTYPING OF COMMON IL28B POLYMORPHISMS: A USEFUL PHARMACOGENETIC TOOL FOR PREDICTION OF HEPATITIS C TREATMENT RESPONSE

Pages

  190-195

Abstract

 Background: In 2009, 3 genome-wide association studies implicated IL28B single-nucleotide POLYMORPHISMs (SNPs) as the strongest GENETIC pretreatment predictor of sustained virological response (SVR) in HEPATITIS C infection. Recently, the American Association for the Study of Liver Diseases (AASLD) and the European Association for the Study of the Liver (EASL) included IL28B testing in their guidelines.Objectives: The main aim of this study was to develop and validate a simple, rapid, and inexpensive polymerase chain reaction-restriction fragment LENGTH POLYMORPHISM (PCR-RFLP) method for genotyping of common IL28B POLYMORPHISMs (rs12979860 and rs8099917).Patients and Methods: Two methods were developed to genotype common IL28B POLYMORPHISMs: 1) PCR-sequencing as a reference method and 2) PCR-RFLP as a rapid and inexpensive method. Both POLYMORPHISMs were genotyped in 104 IRANian HEPATITIS C patients by both methods simultaneously. To validate the PCR-RFLP method, the PCR-RFLP genotyping results should be 100% concordant with the PCR-sequencing results.Results: Genotyping of rs12979860 and rs8099917 by PCR-RFLP was concordant with PCR-sequencing in 104 (100%) individuals. The analytical sensitivity and specificity of the PCR-RFLP method for genotyping of both SNPs are 100%. Among these 104 patients with chronic HEPATITIS C, the frequency of the rs12979860 CC, CT and TT genotypes were 40.4%, 47.1% and 12.5% and the frequency of the rs8099917 TT, GT and GG genotypes were 59.6%, 35.6% and 4.8%, respectively. Also, three IL28B haplotypes (rs12979860-rs8099917) were found among our patients including C-T, T-G and T-T with 63.9%, 22.6% and 13.5% frequency, respectively. C-G haplotype was absent in all of our patients.Conclusions: We have developed a validated, fast, and simple PCR-RFLP method for genotyping of common IL28B SNPs that is more cost-effective than sequencing.

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    APA: Copy

    SHARAFI, HEIDAR, POURYASIN, ALI, ALAVIAN, SEYED MOAYED, BEHNAVA, BITA, KESHVARI, MARYAM, MEHRNOUSH, LEILA, SALIMI, SHIMA, & KHERADVAR, OSVEH. (2012). DEVELOPMENT AND VALIDATION OF A SIMPLE, RAPID AND INEXPENSIVE PCRRFLP METHOD FOR GENOTYPING OF COMMON IL28B POLYMORPHISMS: A USEFUL PHARMACOGENETIC TOOL FOR PREDICTION OF HEPATITIS C TREATMENT RESPONSE. HEPATITIS MONTHLY, 12(3), 190-195. SID. https://sid.ir/paper/306547/en

    Vancouver: Copy

    SHARAFI HEIDAR, POURYASIN ALI, ALAVIAN SEYED MOAYED, BEHNAVA BITA, KESHVARI MARYAM, MEHRNOUSH LEILA, SALIMI SHIMA, KHERADVAR OSVEH. DEVELOPMENT AND VALIDATION OF A SIMPLE, RAPID AND INEXPENSIVE PCRRFLP METHOD FOR GENOTYPING OF COMMON IL28B POLYMORPHISMS: A USEFUL PHARMACOGENETIC TOOL FOR PREDICTION OF HEPATITIS C TREATMENT RESPONSE. HEPATITIS MONTHLY[Internet]. 2012;12(3):190-195. Available from: https://sid.ir/paper/306547/en

    IEEE: Copy

    HEIDAR SHARAFI, ALI POURYASIN, SEYED MOAYED ALAVIAN, BITA BEHNAVA, MARYAM KESHVARI, LEILA MEHRNOUSH, SHIMA SALIMI, and OSVEH KHERADVAR, “DEVELOPMENT AND VALIDATION OF A SIMPLE, RAPID AND INEXPENSIVE PCRRFLP METHOD FOR GENOTYPING OF COMMON IL28B POLYMORPHISMS: A USEFUL PHARMACOGENETIC TOOL FOR PREDICTION OF HEPATITIS C TREATMENT RESPONSE,” HEPATITIS MONTHLY, vol. 12, no. 3, pp. 190–195, 2012, [Online]. Available: https://sid.ir/paper/306547/en

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