scholarly journals Identification of TNIP1 Polymorphisms by High Resolution Melting Analysis with Unlabelled Probe: Association with Systemic Lupus Erythematosus

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Jie Zhang ◽  
Yuewen Chen ◽  
Yong Shao ◽  
Qi Wu ◽  
Ming Guan ◽  
...  
Keyword(s):  
High Resolution ◽  
Lupus Erythematosus ◽  
Chinese Population ◽  
High Resolution Melting ◽  
Disease Risk ◽  
High Resolution Melting Analysis ◽  
Negative Regulator ◽  
Hrm Analysis ◽  
Unlabeled Probe ◽  
Melting Analysis

Background. TNFα-induced protein 3 (TNFAIP3) interacting with protein 1 (TNIP1) acts as a negative regulator of NF-κB and plays an important role in maintaining the homeostasis of immune system. A recent genome-wide association study (GWAS) showed that the polymorphism of TNIP1 was associated with the disease risk of SLE in Caucasian. In this study, we investigated whether the association of TNIP1 with SLE was replicated in Chinese population.Methods. The association of TNIP1 SNP rs7708392 (G/C) was determined by high resolution melting (HRM) analysis with unlabeled probe in 285 SLE patients and 336 healthy controls.Results. A new SNP rs79937737 located on 5 bp upstream of rs7708392 was discovered during the HRM analysis. No association of rs7708392 or rs79937737 with the disease risk of SLE was found. Furthermore, rs7708392 and rs79937737 were in weak linkage disequilibrium (LD). Hypotypes analysis of the two SNPs also showed no association with SLE in Chinese population.Conclusions. High resolution melting analysis with unlabeled probes proves to be a powerful and efficient genotyping method for identifying and screening SNPs. No association of rs7708392 or rs79937737 with the disease risk of SLE was observed in Chinese population.

A High-Resolution Melting Analysis with an Unlabeled Probe for CRISPR/Cas9-Induced ZBED6 Knockout Pigs Detection

2020 ◽  
Author(s):  
Xiaofei Liu ◽  
Songyin Qiu ◽  
Lin Mei ◽  
Hongli Jing ◽  
Xiangmei Lin ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Limit Of Detection ◽  
Economic Traits ◽  
Established Method ◽  
Hrm Analysis ◽  
Unlabeled Probe ◽  
Single Base Deletion ◽  
Melting Analysis ◽  
Tissue Specimens

Abstract Background The zinc finger BED-type containing 6 knockout (ZBED6-KO) pigs were created to improve economic traits by increasing the expression of insulin-like growth factor 2. They were generated by CRISPR/CRISPR-associated protein 9 (Cas9) technology and a single-base deletion of ZBED6 was found. An efficient and rapid method was needed to detect this type of pig. Objective This study aimed to develop a high-resolution melting (HRM) method to detect ZBED6-KO pigs. Methods An unlabeled probe and two primers were designed to develop HRM method. The limit of detection, specificity and accuracy of established method were tested by the constructed plasmid and DNA extracts of tissue specimens. Results The limit of detection by established method was 102 copies/µL. The HRM method with an unlabeled probe showed good specificity and high accuracy. Conclusions The established HRM analysis with an unlabeled probe showed it to be a highly effective, rapid and reliable to distinguish ZBED6-KO pigs with wild-type pigs. Highlights It is the first time that HRM analysis with an unlabeled probe has been used in the detection of genome editing pigs by the CRISPR/Cas9 technology.

Genotyping cytomegalovirus UL97 mutations by high-resolution melting analysis with unlabeled probe

2011 ◽  
Vol 157 (3) ◽  
pp. 475-481 ◽  
Author(s):  
Xiao-Tao Zhao ◽  
Dan-Qiu Zhou ◽  
Shuai Wu ◽  
Yue-Wen Chen ◽  
Yong Shao ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
High Resolution Melting Analysis ◽  
Unlabeled Probe ◽  
Melting Analysis

scholarly journals The Association of IL-12b Polymorphisms with Systemic Lupus Erythematosus in Chinese Han Population

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Yong Shao ◽  
Jie Zhang ◽  
Yuewen Chen ◽  
Qi Wu ◽  
Ming Guan ◽  
...  
Keyword(s):  
Systemic Lupus Erythematosus ◽  
High Resolution ◽  
Lupus Erythematosus ◽  
Antinuclear Antibody ◽  
High Resolution Melting ◽  
Systemic Lupus ◽  
Chinese Han ◽  
Han Population ◽  
Unlabeled Probe ◽  
Melting Analysis

Background. Systemic lupus erythematosus (SLE) is a complex immune disease. The genetic variation in the IL-12b gene was found to associate with SLE in Caucasian population. In this study, we examined this association in Chinese Han population by a recently developed method, unlabeled probe-based high resolution melting analysis.Methods. A total of 297 SLE patients and 351 controls were recruited. Unlabeled probe-based high resolution melting analysis (HRMA) was used in genotyping.Results. Statistically significant differences were observed in both genotype and allele frequencies for rs6887695 in the SLE patients as compared with the controls. Minor allele (C) of rs6887695 (P=0.031, OR 0.78, [95% CI 0.63-0.98]) was found to be protective against SLE. The association of SNP rs6887695 with the diagnostic criteria of SLE was also examined. Minor allele (C) exerts protective effect on the incidence of arthritis (P=0.013, OR = 0.65, 95% CI = 0.47-0.92) and abnormalities of antinuclear antibody (P=0.022, OR = 0.68, 95% CI = 0.49–0.95). IL-12b SNPs were irrelevant to other diagnostic criteria of SLE.Summary. Polymorphisms of rs6887695 in IL-12b gene were associated with disease risk, as well as arthritis and antinuclear antibody synthesis, of systemic lupus erythematosus in Chinese population.

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