scholarly journals ANALYSIS OF GENES IL-1Β (C511T), IL-2 (T330G), IL-13 (C1111T) POLYMORPHISM ASSOCIATION WITH THE DEVELOPMENT OF STOMACH DISEASES

2015 ◽  
pp. 97-102
Author(s):  
A. V. Voropayeva ◽  
O. V. Karpenko ◽  
E. V. Bredikhina ◽  
T. P. Krivun ◽  
O. V. Osipkina ◽  
...  
Keyword(s):  
Gene Polymorphism ◽  
Specific Pcr ◽  
Cancer Of The Stomach ◽  
Genetic Components ◽  
Stomach Diseases ◽  
Pcr Rflp ◽  
The Status ◽  
Allele Specific ◽  
H Pylori ◽  
Allele Specific Pcr

Objective: to study the role of IL-1β (C511T), IL-2 (T330G), IL-13 (C1111T) gene polymorphism in the development of stomach diseases in Belarusian population. Material and methods. The study included 194 patients diagnosed with «chronic gastritis» and 68 patients diagnosedwith «cancer of the stomach». To determine polymorphisms of IL-2 (T330G), IL-13 (C-1111T) and IL-1β (C511T) genes, we used the method of allele-specific PCR (AS-PCR) and PCR detection of the length of restriction fragments (PCR-RFLP). Results . By the loci of IL-2 (T330G), IL-13 (C1111T), IL-1b (S511T) we revealed the correlation with oncopathology depending on the status of infection by various strains of H. pylori. Conclusion . The use of the revealed association of cytokine gene polymorphism with the development of the diseases makes it possible to select genetic components that determine the sensitivityof the population to the development of stomach diseases depending on H. pylori infection in Belarusian population.

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.

Comparison of PCR-RFLP with Allele-Specific PCR in Genetic Testing for Spinal Muscular Atrophy

2003 ◽  
Vol 7 (4) ◽  
pp. 277-281 ◽  
Author(s):  
Ruliang Xu ◽  
Shuji Ogino ◽  
Va Lip ◽  
Hong Fang ◽  
Bai-Lin Wu
Keyword(s):  
Genetic Testing ◽  
Spinal Muscular Atrophy ◽  
Muscular Atrophy ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Allele Specific Pcr

scholarly journals The Efficacy of Laboratory Diagnosis of Helicobacter pylori Infections in Gastric Biopsy Specimens Is Related to Bacterial Density and vacA , cagA , and iceA Genotypes

2000 ◽  
Vol 38 (1) ◽  
pp. 13-17
Author(s):  
Leen-Jan van Doorn ◽  
Yvonne Henskens ◽  
Nathalie Nouhan ◽  
Anita Verschuuren ◽  
Rolf Vreede ◽  
...  
Keyword(s):  
Biopsy Specimen ◽  
Laboratory Diagnosis ◽  
Serum Samples ◽  
Specific Pcr ◽  
Test Culture ◽  
Allele Specific ◽  
Probe Assay ◽  
H Pylori ◽  
Allele Specific Pcr ◽  
Positive Results

ABSTRACT A total of 500 consecutive patients undergoing upper endoscopy were biopsied and tested for H. pylori infection by the Campylobacter -like organism (CLO) test, culture, histology, and PCR. Serum samples were tested by two different serological assays. Patients were considered H. pylori positive if at least two of the four biopsy specimen-based methods yielded positive results. PCR had the highest diagnostic sensitivity (99.4%), followed by histology (92.2%), culture (89.5%), and the CLO test (89.0%). The specificities of all methods were higher than 98%. Of the organisms from the 181 PCR-positive patients, the vacA (s and m regions), cagA , and iceA genotypes were determined by reverse hybridization (line probe assay) or an allele-specific PCR. Organisms that were detected by PCR but that remained undetected by the CLO test were significantly more often vacA s1 ( P = 0.006), m1 ( P = 0.028), and cagA positive ( P = 0.029) than vacA s2, m2, and cagA negative, respectively. Organisms that were detected by PCR but that remained undetected by culture or histology more often contained iceA1 ( P = 0.034 and P = 0.029, respectively) than iceA2 . Higher H. pylori density was associated with vacA s2 ( P = 0.024), vacA m2 ( P = 0.050), and cagA -negative ( P = 0.035) genotypes. Also, the diagnostic results of the CLO test ( P = 0.001) and culture ( P = 0.031) but not those of the PCR ( P = 0.130) were significantly associated with the H. pylori density. The rate of detection by the four biopsy specimen-based tests was lower for patients who used proton pump inhibitors, but this was independent of the H. pylori genotypes. These observations may be explained by different bacterial densities, as established by the distinct genotypes of H. pylori , and confirm that the biologies of strains with such genotypes are considerably different.

A Cross-Sectional Study for Evaluation of KRAS and BRAF Mutations by Reverse Dot Blot, PCR-RFLP, and Allele-Specific PCR Methods Among Patients with Colorectal Cancer

2021 ◽  
Author(s):  
Fatemeh Sheikhsofla ◽  
Behzad Poopak ◽  
Sajjad Firuzyar ◽  
Fatemeh Roudbari ◽  
Mojtaba Ghadiany
Keyword(s):  
Colorectal Cancer ◽  
Gene Mutations ◽  
Dot Blot ◽  
Braf Mutations ◽  
Kras Gene ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Allele Specific Pcr ◽  
Reverse Dot Blot

Background: KRAS and BRAF genes are the biomarkers in Colorectal Cancer (CRC) which play prognostic and predictive roles in CRC treatment. Nowadays, the selection of rapid and available methods for studying KRAS and BRAF mutations in anti-EGFR therapy of patients suffering from CRC plays a significant role. In this study, the mutations of these two oncogenes were evaluated by different methods. Methods: This study was performed on 50 Formalin-Fixed Paraffin-Embedded (FFPE) tissue blocks of patients diagnosed with colorectal cancer. After DNA extraction, KRAS and BRAF gene mutations were evaluated using reverse dot blot, and results were compared with PCR-RFLP and allele-specific PCR for KRAS and BRAF mutations, respectively. Results: KRAS gene mutations were detected in 42% of patients, of which 30% were in codon 12 region, and 12% in codon 13. The most frequent mutations of KRAS were related to G12D  and 10% of patients had BRAF mutated genes. The type of KRAS gene mutations could be evaluated by reverse dot blot method. In general, the results of PCR-RFLP and allele-specific PCR were similar to the findings by reverse dot blot method.  Conclusion: These findings suggest that PCR-RFLP and allele-specific PCR methods are suitable for screening the presence of the mutations in KRAS and BRAF oncogenes. In fact, another method with more sensitivity is needed for a more accurate assessment to determine the type of mutations. Due to higher speed of detection, reduced Turnaround Time (TAT), and possible role of some KRAS point mutations in overall survival, reverse dot blot analysis seems to be an optimal method.

Helicobacter pylori 23S rRNA gene A2142G, A2143G, T2182C, and C2195T mutations associated with clarithromycin resistance detected in Sudanese patients

2020 ◽  
Author(s):  
Aalaa Mahgoub Albasha ◽  
Maram M. Alnosh ◽  
Esraa Hassan Osman ◽  
Duha M Zeinalabdin ◽  
Amira A M Fadl ◽  
...  
Keyword(s):  
Dna Sequencing ◽  
Point Mutations ◽  
23S Rrna ◽  
Rrna Gene ◽  
Specific Pcr ◽  
Clarithromycin Resistance ◽  
23S Rrna Gene ◽  
Allele Specific ◽  
H Pylori ◽  
Allele Specific Pcr

Abstract Background: Clarithromycin resistant Helicobacter pylori (H. pylori) strains represent a worldwide health problem. These stains are usually carrying mutations within the 23S rRNA gene associated with clarithromycin resistance. This study aimed to detect H. pylori and clarithromycin resistant associated mutations from Sudanese patients with gastritis symptoms.Materials and Methods: Two hundred and eighty-eight gastric biopsies were collected using gastrointestinal endoscopy from patients with gastritis symptoms in different hospitals in Khartoum state. H. pylori was detected by PCR using primers targeting 16S rRNA and 23S rRNA. Then allele-specific PCR and DNA sequencing were used to screen for the presence of A2142G and A2143G point mutations.Results: Out of 288 samples, H. pylori was detected in 97 (33.7%) sample. Allele-specific PCR detected the variant A2142G in 9/97 (9.3%) sample, while A2143G mutation was not found in any sample. The DNA sequencing revealed the presence of mutations associated with clarithromycin-resistance in 48% (12/25) of samples; the A2142G was present in one sample, A2143G in 5 samples, T2182C in 4 samples, and C2195T in 3 samples. There was no association of 23S rRNA gene point mutations with gender, age group and geographical distribution of patients.Conclusion: This study revealed a high frequency (48%) of mutations associated with clarithromycin resistance using DNA sequencing of the 23S rRNA gene's V domain. This information should be taken into consideration before choosing optimal therapy for H. pylori eradication.

scholarly journals Helicobacter pylori 23S rRNA gene A2142G, A2143G, T2182C, and C2195T mutations associated with clarithromycin resistance detected in Sudanese patients

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Aalaa Mahgoub Albasha ◽  
Maram M. Elnosh ◽  
Esraa Hassan Osman ◽  
Duha M. Zeinalabdin ◽  
Amira A. M. Fadl ◽  
...  
Keyword(s):  
Dna Sequencing ◽  
Point Mutations ◽  
23S Rrna ◽  
Rrna Gene ◽  
Specific Pcr ◽  
Clarithromycin Resistance ◽  
23S Rrna Gene ◽  
Allele Specific ◽  
H Pylori ◽  
Allele Specific Pcr

Abstract Background Clarithromycin resistant Helicobacter pylori (H. pylori) strains represent a worldwide health problem. These stains are usually carrying mutations within the 23S rRNA gene associated with clarithromycin resistance. This study aimed to detect H. pylori and clarithromycin resistant associated mutations from Sudanese patients with gastritis symptoms. Materials and methods Two hundred and eighty-eight gastric biopsies were collected using gastrointestinal endoscopy from patients with gastritis symptoms in different hospitals in Khartoum state. H. pylori was detected by PCR using primer targeting 16S rRNA. Then allele-specific PCR and DNA sequencing were used to screen for the presence of A2142G and A2143G point mutations. Results Out of 288 samples, H. pylori was detected in 88 (~ 30.6%) samples by 16 s RNA. Allele-specific PCR detected the variant A2142G in 9/53 (~ 17%) sample, while A2143G mutation was not found in any sample. The DNA sequencing revealed the presence of mutations associated with clarithromycin-resistance in 36% (9/25) of samples; the A2142G was present in one sample, A2143G in 5 samples and T2182C in 4 samples. Additionally, another point mutation (C2195T) was detected in 3 samples. There was no association of 23S rRNA gene point mutations with gender, age group, and patients’ geographical distribution. Conclusion This study revealed a high frequency (36%) of mutations associated with clarithromycin resistance using DNA sequencing of the 23S rRNA gene’s V domain. This information should be taken into consideration to avoid eradication therapy failing.

scholarly journals Influence of IL10 (G1082A) and TNFα (G308A) Polymorphisms on the Survival of Pediatric Patients with ALL

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Dayse Maria Vasconcelos de Deus ◽  
Karina Araújo Lugo ◽  
Maria Tereza Cartaxo Muniz
Keyword(s):  
Lymphoblastic Leukemia ◽  
Interleukin 10 ◽  
Promoter Regions ◽  
Necrosis Factor Alpha ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Factor Alpha ◽  
The Impact ◽  
Allele Specific Pcr

Interleukin 10 (IL10) is a pleiotropic cytokine that stimulates various hematopoietic cells. The tumor necrosis factor alpha (TNFα) is a cytokine that may influence the transcriptional activity induced by glucocorticoids. This study examined the impact of TNFα (G308A) and IL10 (G1082A) polymorphisms at promoter regions in relation to the overall survival of 105 children (0≤18 years) with acute lymphoblastic leukemia (ALL) for a period of 126 months, treated according to the protocol GBTLI99. The G1082A and G308A polymorphisms were identified by allele-specific PCR and PCR-RFLP, respectively. Patients with IL10AA genotype had a higher death ratio (44%, P=0.0089). Patients with both IL10AA and TNFAA genotypes showed the worst survival when compared with the IL10GG and TNFGA genotypes (P=0.0043). The results of this study revealed a lower survival among patients with IL10AA genotype and the concomitant occurrence of IL10AA and TNFAA genotypes.

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