Complement C3 Genotyping of Slow and Fast Variants by Real Time PCR-High Resolution Melting

2012 ◽  
Vol 10 (3) ◽  
pp. 329-334 ◽  
Author(s):  
D.M. Valero-Hervás ◽  
P. Morales ◽  
M.J. Castro ◽  
P. Varela ◽  
M. Castillo-Rama ◽  
...  
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Low Cost ◽  
Complement C3 ◽  
Amplification Refractory Mutation System ◽  
Rt Pcr ◽  
Dna Amount ◽  
Mutation System

“Slow” and “Fast” C3 complement variants (C3S and C3F) result from a g.304C>G polymorphism that changes arginine to glycine at position 102. C3 variants are associated with complement-mediated diseases and outcome in transplantation. In this work C3 genotyping is achieved by a Real Time PCR - High Resolution Melting (RT-PCR-HRM) optimized method. In an analysis of 49 subjects, 10.2% were C3FF, 36.7% were C3SF and 53.1% were C3SS. Allelic frequencies (70% for C3S and 30% for C3F) were in Hardy-Weinberg equilibrium and similar to those published previously. When comparing RT-PCR-HRM with the currently used Tetraprimer-Amplification Refractory Mutation System PCR (T-ARMS-PCR), coincidence was 93.8%. The procedure shown here includes a single primer pair and low DNA amount per reaction. Detection of C3 variants by RT-PCR-HRM is accurate, easy, fast and low cost, and it may be the method of choice for C3 genotyping.

DETECTION AND DIFFERENTIATION OF THE VACCINE STRAIN AND FIELD ISOLATES OF DUCK HEPATITIS A VIRUS TYPE 1 USING REAL-TIME RT-PCR AND HIGH RESOLUTION MELTING ASSAYS

2015 ◽  
Vol 41 (04) ◽  
pp. 229-235
Author(s):  
Kuang-Po Li ◽  
Shan-Chia Ou ◽  
Jui-Hung Shien ◽  
Poa-Chun Chang
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Virus Type ◽  
Vaccine Strain ◽  
Hepatitis A ◽  
High Resolution Melting ◽  
Hepatitis A Virus ◽  
Rt Pcr ◽  
Duck Hepatitis

Duck hepatitis A virus type 1 (DHAV-1) infection is a highly contagious and fatal disease of young ducklings. A live attenuated vaccine strain designated as 5886 has been used in Taiwan for the control of DHAV-1. Although several molecular biological methods are reported for diagnosis of DHAV-1 infection, none of them is able to discriminate between the vaccine strain and field viruses of DHAV-1. In the present study, a real-time reverse transcriptase polymerase chain reaction (RT-PCR) and high resolution melting (HRM) assay was developed for rapid detection and differentiation between the vaccine strain and field viruses of DHAV-1. This assay is highly specific for DHAV-1 and the detection limit is about 100 copies of the viral RNA. Experiments using fecal samples collected from ducklings experimentally infected with DHAV-1 showed that DHAV-1 could be detected in fecal samples as early as 6 h post-infection. In summary, a real-time RT-PCR and HRM assay is developed in this study and this assay could be valuable for diagnosis and surveillance of DHAV-1 infection in the field.

Development of a quantitation approach for total human and male DNA based on real time PCR followed by high resolution melting analysis

2016 ◽  
Vol 37 (21) ◽  
pp. 2734-2741 ◽  
Author(s):  
Santiago Ginart ◽  
Mariela Caputo ◽  
Evguenia Alechine ◽  
Daniel Corach ◽  
Andrea Sala
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

Real-time PCR followed by high-resolution melting analysis – a new robust approach to evaluate SCCmectyping of methicillin-resistantStaphylococcus aureus

2019 ◽  
Vol 14 (2) ◽  
pp. 155-164 ◽  
Author(s):  
Yasaman Sadeghi ◽  
Seyed Alireza Salami ◽  
Pegah Kananizadeh ◽  
Sayed-Hamidreza Mozhgani ◽  
Mohammad Reza Pourmand
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
High Resolution Melting Analysis ◽  
Robust Approach ◽  
Melting Analysis

Rapid Molecular Typing of Prototheca zopfii by High Resolution Melting Real-Time PCR (PCR-HRM)

2013 ◽  
Vol 54 (4) ◽  
pp. 341-344 ◽  
Author(s):  
Hideto Sobukawa ◽  
Masato Ibaraki ◽  
Rui Kano ◽  
Takaaki Ito ◽  
Kazuyuki Suzuki ◽  
...  
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
Molecular Typing ◽  
High Resolution Melting ◽  
Prototheca Zopfii
Sign in / Sign up

Export Citation Format

Share Document