Genotyping Interleukin-10 High and Low Producers with Single-Tube Bidirectional Allele-Specific Amplification

2001 ◽  
Vol 18 (2) ◽  
pp. 67-70 ◽  
Author(s):  
Jari Karhukorpi ◽  
Riitta Karttunen
Keyword(s):  
Interleukin 10 ◽  
Single Tube ◽  
Allele Specific ◽  
Specific Amplification

scholarly journals Rapid Detection of the CYP2D6*3, CYP2D6*4, and CYP2D6*6 Alleles by Tetra-Primer PCR and of the CYP2D6*5 Allele by Multiplex Long PCR

2000 ◽  
Vol 46 (8) ◽  
pp. 1072-1077 ◽  
Author(s):  
Martin Hersberger ◽  
Jacqueline Marti-Jaun ◽  
Katharina Rentsch ◽  
Edgar Hänseler
Keyword(s):  
Fragment Length Polymorphism ◽  
Poor Metabolizers ◽  
Pcr Analysis ◽  
Sequencing Analysis ◽  
Cyp2d6 Gene ◽  
Cyp2d6 Activity ◽  
Single Tube ◽  
Long Pcr ◽  
Allele Specific ◽  
Specific Amplification

Abstract Background: Interindividual differences in CYP2D6 activity range from total absence of metabolism of certain drugs to ultrafast metabolism and can produce adverse effects or lack of therapeutic effect under standard therapy. Several mutations have been described in the CYP2D6 gene that abolish CYP2D6 activity. However, four mutations explain the majority of the poor metabolizers. We describe four single-tube assays to detect these mutations. Methods: Three tetra-primer PCR assays were developed to detect the mutations in the CYP2D6*3, *4, and *6 alleles. In these single-tube assays, the CYP2D6 locus is amplified directly, followed by the allele-specific amplification on this new template. In addition, a multiplex long PCR was developed to genotype the CYP2D6*5 allele. Two long PCR amplifications for detection of the deletion of CYP2D6 (*5) and for detection of the CYP2D6 gene region were combined in one tube. Results: Analysis of 114 alleles showed no CYP2D6*3 allele, and allele frequencies of 28.1% for CYP2D6*4, 2.6% for CYP2D6*5, and 0.9% for CYP2D6*6. Re-analysis of the DNA samples by restriction fragment length polymorphism and sequencing analysis confirmed these results. Furthermore, re-analysis of sequenced genomic DNA by tetra-primer PCR analysis (7–11 times) always showed identical results. Conclusions: Our set of single-tube assays allows rapid and reproducible genotyping of the majority of CYP2D6 poor metabolizers.

Simple and Rapid Detection of Factor V Leiden by Allele-specific PCR Amplification

1996 ◽  
Vol 75 (05) ◽  
pp. 757-759 ◽  
Author(s):  
Rainer Blasczyk ◽  
Markus Ritter ◽  
Christian Thiede ◽  
Jenny Wehling ◽  
Günter Hintz ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Pcr Amplification ◽  
Factor V ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Specific Amplification ◽  
Allele Specific Pcr

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.

Individual single tube genotyping and DNA pooling by allele-specific PCR to uncover associations of polymorphisms with complex diseases

2007 ◽  
Vol 376 (1-2) ◽  
pp. 155-162 ◽  
Author(s):  
Antonio Casado-Díaz ◽  
Rafael Cuenca-Acevedo ◽  
José Manuel Quesada ◽  
Gabriel Dorado
Keyword(s):  
Complex Diseases ◽  
Dna Pooling ◽  
Single Tube ◽  
Specific Pcr ◽  
Allele Specific ◽  
Allele Specific Pcr

scholarly journals Detection of rare point mutation via allele-specific amplification in emulsion PCR

BMB Reports ◽  
2013 ◽  
Vol 46 (5) ◽  
pp. 270-275 ◽  
Author(s):  
Changming Cheng ◽  
Yin Zhou ◽  
Chao Yang ◽  
Juan Chen ◽  
Jie Wang ◽  
...  
Keyword(s):  
Point Mutation ◽  
Emulsion Pcr ◽  
Allele Specific ◽  
Specific Amplification

scholarly journals Epidermal Growth Factor rs4444903 A>G Gene Polymorphism Association with Chronic Hepatitis B Liver Disease Progression among Egyptian Children: A Multicenter Study

2021 ◽  
Vol 11 (1) ◽  
pp. 63-68
Author(s):  
Amal A. Mohamed ◽  
Gehan L.A. Hakeem ◽  
Gihan M. Babrs ◽  
Laila E. Abolfotoh ◽  
Nageh M. Shehata ◽  
...  
Keyword(s):  
Chronic Hepatitis ◽  
Hepatitis B ◽  
Chronic Hepatitis B ◽  
Genotypic Frequency ◽  
Liver Disease Progression ◽  
Egyptian Children ◽  
Chronic Hepatitis B Virus ◽  
Genes Encoding ◽  
Allele Specific ◽  
Specific Amplification

Background: Polymorphisms of genes encoding the pro-inflammatory and anti-inflammatory cytokines can affect the clinical presentation of the infection. We aimed to assess the role of EGF gene single-nucleotide polymorphism in the outcome of chronic hepatitis B virus (HBV) infection in children. Methods: One hundred HBV-infected children and 75 healthy matched controls were enrolled in this prospective study. Patients included 18 chronic inactive and 82 chronic active carriers. EGF rs4444903 A>G genotypes were determined using allele-specific amplification. Results: Significant differences regarding EGF genotypic frequency (p=0.001) in patients compared to controls (p=0.001). Eighteen percent were inactive, and 82% were active carriers. AA, AG and GG genotypic frequency were 66.7%, 33.3%, 0% and were 3.7%, 37.8% and 58.5% in the inactive and active carriers, respectively, with significant differences regarding AA, AG, GG genotypic frequency (p=0.001 for all). EGF AA, AG, GG genotypes frequency were 1.9%, 33.3%, and 64.8%, respectively, with significant differences between cirrhotic and non-cirrhotic patients regarding AA, AG, GG genotypic frequency (p=0.001 for all). Conclusion: Increased G allele frequency in EGF rs4444903 A > G polymorphism in HBV- Egyptian children is associated with worse outcomes.

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