title: Investigation of sensitivity, specificity and accuracy of Tetra primer ARMS PCR method in comparison with conventional ARMS PCR, based on sequencing technique outcomes in IVS-II-I genotyping of beta thalassemia patients.
abstract: OBJECTIVE
  Beta thalassemia is one of the most important hematic diseases all around the world and solving the problems caused by this abnormality is strongly dependent on precise detection and reliable screening of high-risk couples. The aim of our study was the investigation of sensitivity, specificity and accuracy of Tetra primer ARMS PCR method comparing with conventional ARMS PCR, based on sequencing technique outcomes for genotyping of IVS-II-I mutation in beta thalassemia patients.
  
  
  METHODS
  Fifty seven samples including two homozygote, 49 heterozygote and 6 normal specimens were analyzed by Tetra primer ARMS PCR and conventional ARMS PCR methods. DNA was extracted by the standard method of salting out for leukocyte genomic DNA extraction of blood specimens and a high pure PCR template preparation kit was used for DNA purification of CVS samples. The results obtained by Tetra primer ARMS PCR and conventional ARMS PCR methods were compared with gold standard technique, i.e. sequencing.
  
  
  RESULTS
  All three parameters including specificity, sensitivity and accuracy were 100% for Tetra primer ARMS PCR method, while they were 100%, 92.45% and 92.7% for conventional ARMS PCR technique respectively. Comparing with Tetra primer ARMS PCR which represented 100% agreement with sequencing method, conventional ARMS PCR technique only showed 47.1% agreement, because of 4 discordant results.
  
  
  CONCLUSIONS
  Tetra primer ARMS PCR method is an almost reliable, sensitive and accurate technique and it is suggested that it can be used as a complementary method for diagnostic cases instead of conventional ARMS PCR method. This suggestion originated with perfect rate of agreement between outcomes of sequencing method, as a gold standard method of detecting the mutations, and Tetra primer ARMS PCR technique comparing with conventional ARMS PCR method.
authors.affiliation: Division of Cellular and Molecular Biology, Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Electronic address: mo.honardost@yahoo.com.
authors.forename: Mohammad Amin
authors.initials: MA
authors.lastname: Honardoost
authors.affiliation: Genetics Division, Biology Department, Faculty of Sciences, University of Isfahan, Isfahan, Iran. Electronic address: Hosein.tabatabaeian@gmail.com.
authors.forename: Hosein
authors.initials: H
authors.lastname: Tabatabaeian
authors.affiliation: School of Medicine, Isfahan University of Medical Science, Isfahan, Iran. Electronic address: m_akbari@med.mui.ac.ir.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Akbari
authors.affiliation: School of Medicine, Isfahan University of Medical Science, Isfahan, Iran; Medical Genetics Center of Genome, Shariati St., Isfahan, Iran. Electronic address: m_salehi@med.mui.ac.ir.
authors.forename: Mansoor
authors.initials: M
authors.lastname: Salehi
mesh_terms: D015193:Chorionic Villi Sampling; D017931:DNA Primers; D005260:Female; D005838:Genotype; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008297:Male; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D055544:beta-Globins / Q000235:genetics*; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics
chemical_list: D017931:DNA Primers; D055544:beta-Globins
keywords: Beta thalassemia; Conventional ARMS PCR; Sequencing; Tetra primer ARMS PCR