Molecular diagnosis of β‐thalassemia by multiplex ARMS‐PCR: a cost effective method for developing countries like Pakistan

Abstract

A MEL‐type pure‐silica zeolite (PSZ), prepared by spin‐on of nanoparticle suspensions, has been shown to be a promising ultra‐low‐dielectric‐constant (k) material because of its high mechanical strength, hydrophobicity, and chemical stability. In our previous works, a two‐stage synthesis method was used to synthesize a MEL‐zeolite nanoparticle suspension, in which both nanocrystal yield and particle size of the zeolite suspension increased with increasing synthesis time. For instance, at a crystal yield of 63%, the particle size is 80 nm, which has proved to be too large because it introduces a number of problems for the spin‐on films, including large surface roughness, surface striations, and large mesopores. In the current study, the two‐stage synthesis method is modified into an evaporation‐assisted two‐stage method by adding a solvent‐evaporation process between the two thermal‐treatment steps. The modified method can yield much smaller particle sizes (e.g., 14 vs. 80 nm) while maintaining the same nanocrystal yields as the two‐stage synthesis. Furthermore, the nanoparticle suspensions from the evaporation‐assisted two‐stage synthesis show a bimodal particle size distribution. The primary nanoparticles are around 14 nm in size and are stable in the final suspension with 60% solvent evaporation. The factors that affect nanocrystal synthesis are discussed, including the concentration, pH value, and viscosity. Spin‐on films prepared by using suspensions synthesized this way have no striations and improved elastic modulus (9.67 ± 1.48 GPa vs. 7.82 ± 1.30 GPa), as well as a similar k value (1.91 ± 0.09 vs. 1.89 ± 0.08) to the previous two‐stage synthesized films.

β-Thalassemia in Pakistan

“…13 Monoplex and/or Multiplex ARMS-PCR was used for detection of known mutations. 14,15 In addition to amplification of internal control region in each reaction, homozygote, heterozygote, and normal control samples for each b-thalassemia mutation screened were included with all ARMS-PCR assays to avoid false-negative and false-positive results. The BigDye Terminator v1.1 Cycle Sequencing Kit (Applied Biosystems) was used in the ABI 3730 DNA analyzer (Applied Biosystems) to sequence coding and noncoding regions (all 3 exons and 2 introns) of b-globin gene to confirm ambiguous results and detection of less common or rare mutations.…”

An Early Diagnosis of Thalassemia: A Boon to a Healthy Society

“…The ARMS primers have been made for the detection of common mutations of β-thalassemia [31]. In various countries like India and Pakistan, this technique quit well-known for screening and prenatal diagnosis due to its quick and low cost [32,33]. The ARMS PCR is able to detect multiple known mutations in a single assay [29,34,35].…”

Prevention of β-Thalassemia in a Large Pakistani Family through Cascade Testing

“…β-Thalassemia was diagnosed at a Hb A 2 of 3.5% or more. Amplification refractory mutation system polymerase chain reaction was used to identify the specific mutation [11, 12]. …”