High-resolution melting analysis for noninvasive prenatal diagnosis of IVS-II-I (G-A) fetal DNA in minor beta-thalassemia mothers

Abstract

HRM real-time PCR was a sensitive and specific method for determining the paternally inherited mutation in the fetus at risk with thalassemia major.

Optimal non‐invasive diagnosis of fetal achondroplasia combining ultrasonography with circulating cell‐free fetal DNA analysis

“…All results were consistent with the genotypes obtained using conventional techniques, while HRM‐PCR reduced the time needed from 3 days to 1 day. These results were recently confirmed by Zafari et al …”

“…All results were consistent with the genotypes obtained using conventional techniques, while HRM-PCR reduced the time needed from 3 days to 1 day. These results were recently confirmed by Zafari et al 19 . Combined with co-amplification at lower denaturation-temperature PCR, HRM is more sensitive than is conventional PCR for the detection of very-low-level mutations 24 and has been applied with very promising results to non-invasive determination of the fetal HPA-1 genotype by Ferro et al 18 in the context of fetal-maternal platelet incompatibility.…”

“…HRM polymerase chain reaction (HRM‐PCR) is an efficient and sensitive tool for the detection of mutations, not only in oncology but also in NIPT. We therefore decided to apply this strategy for the first time to achondroplasia.…”

Allele-Specific Loop-Mediated Isothermal Amplification for the Detection of IVSII-I G>A Mutation On β-Globin Gene

“…The purpose of the research was to detect the fetus’s IVSII-I G◊A mutation in the serum of pregnant mothers. The sensitivity and specificity of the introduced method were 92.6% and 82.6%, respectively [32].…”

Fluorescent detection of point mutation via ligase reaction assisted by quantum dots and magnetic nanoparticle-based probes

“…Several point mutation detection methods have been developed such as restriction fragment length polymorphisms (RFLP), 52 temporal temperature gradient gel electrophoresis (TGGE), 53 denaturing gradient gel electrophoresis (DGGE), 54,55 reverse dot blot hybridization, amplication refractory mutation system (ARMS), 56 high resolution melting (HRM). 35 However, these technologies have some shortcomings. The main disadvantages of PCR based methods although highly sensitive are amplication errors due to mispriming, limited accuracy of discriminating single nucleotide variations, post-PCR processing steps such as gel electrophoresis and limited multiplexing capability.…”