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A facile PCR-RFLP method for genotyping of ITPA rs1127354 and rs7270101 polymorphisms.
Alavian, Seyed Ehsan; Sharafi, Heidar; Shirmast, Paniz; Alavian, Seyed Moayed; Behnava, Bita; Pouryasin, Mohammad; Keshvari, Maryam; Pouryasin, Ali.
Afiliação
  • Alavian SE; Baqiyatallah Research Center for Gastroenterology and Liver Diseases, Baqiyatallah University of Medical Sciences, Tehran, Iran.
  • Sharafi H; Middle East Liver Diseases (MELD) Virology Laboratory, MELD Center, Tehran, Iran.
  • Shirmast P; Baqiyatallah Research Center for Gastroenterology and Liver Diseases, Baqiyatallah University of Medical Sciences, Tehran, Iran.
  • Alavian SM; Middle East Liver Diseases (MELD) Virology Laboratory, MELD Center, Tehran, Iran.
  • Behnava B; Middle East Liver Diseases (MELD) Virology Laboratory, MELD Center, Tehran, Iran.
  • Pouryasin M; Baqiyatallah Research Center for Gastroenterology and Liver Diseases, Baqiyatallah University of Medical Sciences, Tehran, Iran.
  • Keshvari M; Baqiyatallah Research Center for Gastroenterology and Liver Diseases, Baqiyatallah University of Medical Sciences, Tehran, Iran.
  • Pouryasin A; Armin Pathobiology Laboratory, Tehran, Iran.
J Clin Lab Anal ; 32(7): e22440, 2018 Sep.
Article em En | MEDLINE | ID: mdl-29660762
ABSTRACT

BACKGROUND:

Inosine triphosphate pyrophosphatase (ITPA) gene single nucleotide polymorphisms (SNPs), rs1127354 and rs7270101, may cause a functional impairment in ITPase enzyme, resulting anemia protection in patients with chronic hepatitis C virus (HCV) infection undergoing ribavirin (RBV)-dependent regimens. The main purpose of this study was to provide and validate a simple, rapid, and inexpensive polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique for genotyping of ITPA rs1127354 and rs7270101 polymorphisms in chronic HCV-infected patients.

METHODS:

In the current study, 100 Iranian patients with chronic hepatitis C were examined and genotyped for ITPA rs1127354 and rs7270101 gene polymorphisms. To genotype rs1127354 and rs7270101 polymorphisms, PCR-RFLP technique and sequencing technique were performed on these samples. To validate the PCR-RFLP method, the PCR-RFLP genotyping results should be 100% concordant with the PCR-sequencing results.

RESULTS:

The rs1127354 and rs7270101 polymorphisms of ITPA gene were genotyped by PCR-RFLP technique and sequencing simultaneously, and the results of both techniques were 100% concordant in all 100 patients. Both PCR-RFLP and sequencing techniques indicated that the genotypic frequency of rs7270101 was 80% AA, 19% AC and 1% CC, and for rs1127354 was 79% CC, 20% CA and 1% AA, respectively.

CONCLUSION:

We developed and validated a rapid and inexpensive PCR-RFLP technique for the detection of ITPA rs1127354 and rs7270101 gene polymorphisms.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pirofosfatases / Polimorfismo de Fragmento de Restrição / Hepatite C Crônica / Técnicas de Genotipagem Tipo de estudo: Etiology_studies / Incidence_studies / Observational_studies / Risk_factors_studies Limite: Female / Humans / Male País/Região como assunto: Asia Idioma: En Revista: J Clin Lab Anal Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Irã

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pirofosfatases / Polimorfismo de Fragmento de Restrição / Hepatite C Crônica / Técnicas de Genotipagem Tipo de estudo: Etiology_studies / Incidence_studies / Observational_studies / Risk_factors_studies Limite: Female / Humans / Male País/Região como assunto: Asia Idioma: En Revista: J Clin Lab Anal Assunto da revista: TECNICAS E PROCEDIMENTOS DE LABORATORIO Ano de publicação: 2018 Tipo de documento: Article País de afiliação: Irã