What is PCR test for thalassemia?
Polymerase chain reaction (PCR) diagnosis of thalassemia usually relies on using genomic DNA. Preparing the genomic DNA can lead to sample-to-sample contamination. This report was aimed to establish the PCR protocol using whole-blood for detecting mutations of α- and β-globin genes causing the thalassemia syndrome.
What is ARMS-PCR?
The Amplification Refractory Mutation System PCR (ARMS-PCR) is one of the most accurate tools in genetic disease diagnosis in recent days. It is a gold standard method for thalassemia and sickle cell anemia. ARMS-PCR is used for genotyping SNP (single nucleotide polymorphism) with the help of refractory primers.
Which of the following PCR techniques is best suited for the detection of a point mutation in DNA?
Amplification refractory mutation system (ARMS) PCR: Allele-specific amplification (AS-PCR) or ARMS-PCR is a general technique for the detection of any point mutation or small deletion.
How does allele specific PCR work?
The Allele-specific PCR has the power to detect a single specific allele. Meaning, If you wish to amplify only a mutant allele, design a primer set accordingly and amplify it using this technique. Each set of specific primers is designed for each specific allele.
What are arms primers?
The amplification-refractory mutation system (ARMS) is a simple method for detecting any mutation involving single base changes or small deletions. ARMS is based on the use of sequence-specific PCR primers that allow amplification of test DNA only when the target allele is contained within the sample.
What is Tetra primer ARMS PCR?
Abstract. The tetra-primer amplification refractory mutation system-polymerase chain (ARMS-PCR) reaction is a simple and economical method to genotype single-nucleotide polymorphisms (SNPs). It uses four primers in a single PCR and is followed just by gel electrophoresis.
What is the difference between a Singleplex and a multiplex PCR?
In conventional singleplex PCR, a single target is amplified in a single reaction tube. In contrast, multiplex PCR allows for simultaneous amplification of multiple target sequences in a single tube using specific primer sets in combination with probes labeled with spectrally distinct fluorophores.
How many primers are in multiplex PCR?
In multiplex PCR, two or more primer sets designed for amplification of different targets are included in the same PCR reaction.
How is PCR used to diagnose β thalassemia?
Most of the molecular diagnostic laboratories utilize PCR amplification, with allele-specific amplification using an amplification refractory mutation system (ARMS), allele-specific oligonucleotide probes (ASO), or reverse dot blot methodology to define the known mutations that cause β-thalassemia [14], [15], [16].
Which is the most common mutation in β-thalassemia?
Several high prevalence β-Thalassemia trait groups constituted by Muslims, Patels, Sindhis, ModhBanias, and Mahayavanshi. Four most common mutations detected in them are IVS I-5 (G→C), Codon 41/42 (- TCTT), 619-bp deletion and FS 8/9 (+G). We identified each of these β-thalassemia mutations in multiplexed ARMS from positive control samples.
Is it possible to diagnose β-thalassaemia in Surat?
Heterogeneity of β-thalassaemia mutations in multi-ethnic population of Surat, makes molecular diagnosis expensive and time consuming.
How are specific primers used in multiplex ARMS PCR?
Specific primers were used to differentiate four common mutations, IVS I-5 (G→C), Codon 41/42 (- TCTT), 619-bp deletion and FS 8/9 (+G), by a simple PCR involving a multiplex amplification refractory mutation system. Several high prevalence β-Thalassemia trait groups constituted by Muslims, Patels, Sindhis, ModhBanias, and Mahayavanshi.