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CURATOR

PhD scholar and lecturer, LIAQUAT UNIVERSITY OF MEDICAL AND HEALTH SCIENCES JAMSHORO

PINBOARD SUMMARY

Comparison of three methods for genotyping

FVL and FV A6755G polymorphisms have been studied in various population previoulsy. However their prevalance in pakistani population has not been determined in girls of reproductive age. As these polymorphisms refer to thrombophilic polymorphisms thier occurance in population will help to determine thier role in certain thrombophilic disorders. Moreover in developing country like Pakistan a cost effective method for genotyping must be estabilished so that various polymorphisms associated with disorders may be deteemined. In our study we have selected late adolescent girls and genotyping was carried out for these polymorphisms and three methods that is PCR RFLP, Tetra ARMS PCR and direct sequencing were compared for their validity, reproducibilty, cost effectiveness and rapidity to estabilish effective genotyping method in low economic setup

1 ITEMS PINNED

Evaluation of amplification refractory mutation system (ARMS) technique for quick and accurate prenatal gene diagnosis of CHM variant in choroideremia.

Abstract: Choroideremia is a rare X-linked recessive inherited disorder that causes chorioretinal dystrophy leading to visual impairment in its early stages which finally causes total blindness in the affected person. It is caused due to mutations in the CHM gene. In this study, we have recruited a pedigree with choroideremia and detected a nonsense variant (c.C799T:p.R267X) in CHM of the proband (I:1). Different primer sets for amplification refractory mutation system (ARMS) were designed and PCR conditions were optimized. Then, we evaluated the sequence variant in the patient, carrier, and a fetus by using ARMS technique to identify if they inherited the pathogenic gene from parental generation; we used amniotic fluid DNA for the diagnosis of the gene in the fetus. The primer pairs, WT2+C and MT+C, amplified high specific products in different DNAs which were verified by Sanger sequencing. Based on our results, ARMS technique is fast, accurate, and reliable prenatal gene diagnostic tool to assess CHM variants. Taken together, our study indicates that ARMS technique can be used as a potential molecular tool in the diagnosis of prenatal mutation for choroideremia as well as other genetic diseases in undeveloped and developing countries, where there might be shortage of medical resources and supplies.

Pub.: 04 Jan '18, Pinned: 15 Jan '18