scholarly journals Molecular Study of Benzimidazole Resistance in Teladorsagia circumcincta Isolated from Sheep in North of Iran

2019 ◽  
Author(s):  
Rahim NEMATI ◽  
Aliasghar BAHARI ◽  
Pezhman MAHMOODI ◽  
Alireza SAZMAND
Keyword(s):  
Anthelmintic Resistance ◽  
Molecular Study ◽  
Nucleotide Polymorphisms ◽  
Northern Iran ◽  
The North ◽  
Specific Pcr ◽  
Teladorsagia Circumcincta ◽  
Pcr Rflp ◽  
North Of Iran ◽  
Allele Specific

Background: Resistance to benzimidazole (BZ) compounds is common in Teladorsagia circumcincta populations in sheep and goats worldwide. Given the importance of anthelmintic resistance and shortage of information on single nucleotide polymorphisms (SNPs) in this prevalent nematode in Iran, this study was conducted. Methods: From June to September 2016, abomasa of 139 sheep of different sexes and ages in Amol City slaughterhouse, northern Iran were examined for isolation of nematodes. Totally 45 male T. circumcincta confirmed by both microscopical and nested-PCR-RFLP methods were included in this study. Susceptibility or resistance of each single T. circumcincta worm to benzimidazoles was assessed using allele-specific PCR. Results: Frequency of genotypes in the present study were 33.33% heterozygote BZ and 66.67% BZ homozygote sensitive. No homozygote resistant worm was found. Conclusion: Resistance against BZs in T. circumcincta of sheep has occurred at a low prevalence in the north of Iran. However, mutated genes might get dominant under drug selection in future. Hence, periodic investigations for early detection of mutated alleles in nematode populations using accurate and sensitive molecular methods such as PCR-RFLP is recommended.

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.

scholarly journals Fine Mapping of qd1, a Dominant Gene that Regulates Stem Elongation in Bread Wheat

2021 ◽  
Vol 12 ◽  
Author(s):  
Yongdun Xie ◽  
Weiwei Zeng ◽  
Chaojie Wang ◽  
Daxing Xu ◽  
Huijun Guo ◽  
...  
Keyword(s):  
Bread Wheat ◽  
Stem Elongation ◽  
Dominant Gene ◽  
Nucleotide Polymorphisms ◽  
Sequencing Data ◽  
Specific Pcr ◽  
Yield Determination ◽  
Allele Specific ◽  
Stem Development ◽  
Kasp Markers

Stem elongation is a critical phase for yield determination and, as a major trait, is targeted for manipulation for improvement in bread wheat (Triticum aestivum L.). In a previous study, we characterized a mutant showing rapid stem elongation but with no effect on plant height at maturity. The present study aimed to finely map the underlying mutated gene, qd1, in this mutant. By analyzing an F2 segregating population consisting of 606 individuals, we found that the qd1 gene behaved in a dominant manner. Moreover, by using the bulked segregant RNA sequencing (BSR-seq)-based linkage analysis method, we initially mapped the qd1 gene to a 13.55 Mb region on chromosome 4B (from 15.41 to 28.96 Mb). This result was further confirmed in F2 and BC3F2 segregating populations. Furthermore, by using transcriptome sequencing data, we developed 14 Kompetitive Allele-Specific PCR (KASP) markers and then mapped the qd1 gene to a smaller and more precise 5.08 Mb interval from 26.80 to 31.88 Mb. To develop additional markers to finely map the qd1 gene, a total of 4,481 single-nucleotide polymorphisms (SNPs) within the 5.08 Mb interval were screened, and 25 KASP markers were developed based on 10x-depth genome resequencing data from both wild-type (WT) and mutant plants. The qd1 gene was finally mapped to a 1.33 Mb interval from 28.86 to 30.19 Mb on chromosome 4B. Four candidate genes were identified in this region. Among them, the expression pattern of only TraesCS4B02G042300 in the stems was concurrent with the stem development of the mutant and WT. The qd1 gene could be used in conjunction with molecular markers to manipulate stem development in the future.

The significance of switch in convergence direction in the Alborz Mountains, northern Iran: Insights from scaled analogue modeling

2017 ◽  
Vol 5 (1) ◽  
pp. SD81-SD98 ◽  
Author(s):  
Majid Shahpasandzadeh ◽  
Hemin Koyi ◽  
Faramarz Nilfouroushan
Keyword(s):  
Active Tectonics ◽  
Deformation Pattern ◽  
Eurasian Plate ◽  
The South ◽  
South Caspian Basin ◽  
Northern Iran ◽  
The North ◽  
Structural Style ◽  
Analogue Models ◽  
North Of Iran

The switch in direction of convergence between Central Iran and the Eurasian Plate is believed to have a significant impact on the structural style in the Alborz Mountains, in the north of Iran. To understand the deformation pattern and investigate the influence of the South Caspian Basin kinematics since the middle Miocene on the structural styles and active tectonics of the Alborz Mountains, a series of scaled analogue models were prepared, in which passively layered loose sand simulating the sedimentary units were subjected to orthogonal and subsequently oblique shortening by a rigid indenter. Model results indicate that during the shortening, an arcuate-shaped foreland-vergent imbricate stack forms in front of the indenter. The orthogonal shortening is characterized by a prevailing right-lateral and left-lateral oblique-slip motion in the east and west of the model, respectively. This shift in kinematics contradicts the proposed preneotectonic (orthogonal) model of the Alborz. However, during oblique shortening, model results show that deformation is mainly accommodated by left-lateral transpression within the sand wedge and internal deformation. Oblique shortening is consistently accommodated by continued left-lateral motion on the west-northwest-trending oblique thrusts, whereas the east–west-trending thrusts and the preexisting east-northeast-trending right-lateral oblique thrusts reactivate as left-lateral oblique faults. Precise monitoring of the model surface also illustrates partitioning of shortening into the foreland-vergent left-lateral thrusting in the south and hinterland-vergent back thrusting in the north. These model results are generally consistent with field observations and GPS data of structure and kinematics of the Alborz Mountains.

scholarly journals Preventing Favism by Selecting Faba Bean Mutants Using Molecular Markers

2017 ◽  
Vol 3 (1) ◽  
pp. 2-6 ◽  
Author(s):  
Melody Song
Keyword(s):  
Faba Bean ◽  
Expressed Sequence Tag ◽  
Nucleotide Polymorphisms ◽  
Seed Selection ◽  
Single Nucleotide ◽  
Specific Pcr ◽  
Faba Beans ◽  
Allele Specific ◽  
Haemolytic Anemia ◽  
Allele Specific Pcr

Faba bean (Vicia faba) is an ancient legume species known for its high protein content. The usage and consumption of the faba bean is limited by a glycoside, vicine-convicine (VC). Consumption of VC causes haemolytic anemia in individuals with the genetic condition called favism. Faba beans with low VC concentration are opening the possibility of reduction of favism disease, but there are many challenges in analyzing VC concentration. The objective of this study was to develop expressed sequence tag (EST) markers that can differentiate between low VC content (LVC) and high VC content (HVC) faba bean genotypes. Three single nucleotide polymorphisms (SNPs) were discovered that distinguished between LVC and HVC genotypes. The SNPs were validated using Kompetitive Allele Specific PCR (KASP) and mass spectrometry phenotyping. Molecular marker SNP 316 (Intron of Medtr2g009270 at 1,851,012 bp) was the most successful marker in differentiating between LVC, HVC, and heterozygous faba bean genotypes. This marker has applications in seed selection and acceleration of breeding programs, which is the first step towards allowing all consumers concerned with the effects of favism to enjoy the nutritional value of faba bean.

scholarly journals Association of Mycobacterium avium subsp. paratuberculosis and SLC11A1 polymorphisms in Sardinian multiple sclerosis patients

2013 ◽  
Vol 7 (03) ◽  
pp. 203-207 ◽  
Author(s):  
Davide Cossu ◽  
Speranza Masala ◽  
Eleonora Cocco ◽  
Daniela Paccagnini ◽  
Stefania Tranquilli ◽  
...  
Keyword(s):  
Multiple Sclerosis ◽  
Autoimmune Disease ◽  
Mycobacterium Avium ◽  
Humoral Response ◽  
Mycobacterium Avium Subsp ◽  
Nucleotide Polymorphisms ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Multiple Sclerosis Patients

Introduction: Recent findings propose that Mycobacterium avium subsp. paratuberculosis (MAP) infection could act as risk factor in favoring multiple sclerosis (MS) progression. SLC11A1 is a gene associated with mycobacterial survival in the host and it may be involved in the induction and maintenance of autoimmune disease. Methodology: In this preliminary study, 100 MS patients and 100 healthy controls (HCs) from Sardinia were enrolled. Eight single nucleotide polymorphisms (SNPs) in the SLC11A gene were searched by PCR RFLP-genotyping. IS900 specie specific PCR was undertaken to search for MAP presence. Indirect ELISA was performed to asses if MS patients displayed a stronger humoral response against MAP2694 protein compared to the HCs. Results: Only rs2276631 SNP was associated with MS. MAP DNA was detected in 23 out of 100 MS patients (23%) and in 7 out of 100 HCs (7%). A strong humoral response against MAP2694 protein was detected in 36% of MS patients and only in 3% of HCs. A correlation between ELISA sero-positivity and the rs2276631 SNP was also found. Conclusion:  Our preliminary results suggest that the Sardinian population might be prone to develop autoimmune disease due to polymorphisms in immunomodulating the SLC11A1 gene, which is important in the immune response against intracellular bacteria such as MAP.

scholarly journals High frequency of benzimidazole resistance alleles in Haemonchus contortus, Teladorsagia circumcincta and Trichostrongylus colubriformis from Austrian sheep flocks in an alpine transhumance management system

2019 ◽  
Author(s):  
Barbara Hinney ◽  
Julia Schoiswohl ◽  
Lynsey Melville ◽  
Vahel J. Ameen ◽  
Walpurga Wille-Piazzai ◽  
...  
Keyword(s):  
Management System ◽  
Anthelmintic Resistance ◽  
Treatment Strategies ◽  
Small Ruminants ◽  
Nucleotide Polymorphisms ◽  
Sheep Population ◽  
Teladorsagia Circumcincta ◽  
Benzimidazole Resistance ◽  
Drug Classes ◽  
New Treatment

Abstract Infections of small ruminants with trichostrongyloid nematodes often result in reduced productivity and may be detrimental to the host. Anthelmintic resistance (AR) against most anthelmintic drug classes is now widespread amongst the trichostrongyloids. Baseline establishment, followed by regular monitoring of the level of AR, is necessary for farmers and veterinarians to make informed decisions about parasite management. The detection of single nucleotide polymorphisms (SNPs) is a sensitive method to detect AR against benzimidazoles (BZs), one of the most widely used anthelmintic classes. Alpine transhumance constitutes a special type of pasturing of sheep from many different farms, the aim of this study was to investigate the prevalence of benzimidazole resistance alleles in this particular management system. Sixteen sheep flocks in Styria and Salzburg in Austria were examined by pyrosequencing for SNPs at codons 167, 198 and 200 of the isotype-1 β-tubulin gene. The frequency of the resistance-associated mutation F200Y was 87–100% for H. contortus, 77–100% for T. colubriformis and <5–66% for T. circumcincta. Additionally, the F167Y polymorphism was detected in T. colubriformis from two farms at a frequency of 19% and 23% respectively. The high resistance allele frequency in H. contortus and T. colubriformis in the examined sheep population urgently calls for the development of new treatment strategies to sustainably control trichostrongyloid infections for this kind of pasturing, since the frequent mixing of flocks during the alpine summer grazing must be considered an important risk factor for the spread of resistant nematodes to a large number of farms.

scholarly journals TNF-α -308G/A polymorphism and susceptibility to tuberculosis in Azeri population of Iran

Genetika ◽  
2016 ◽  
Vol 48 (3) ◽  
pp. 819-826 ◽  
Author(s):  
Sajjad Ghorghanlu ◽  
Mohammad Asgharzadeh ◽  
Hossein Samadi-Kafil ◽  
Fatemeh Khaki-Khatibi ◽  
Jalil Rashedi ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Tuberculosis Patient ◽  
P Value ◽  
Nucleotide Polymorphisms ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Tuberculosis Patients ◽  
Specific Pcr ◽  
Allele Specific ◽  
And Function

Single nucleotide polymorphisms (SNPs) in cytokine genes may alter the level and function of secreted cytokine; therefore, SNPs can influence the immune response. The aim of the present study was to determine the association of TNF-? -308G/A single nucleotide polymorphism in tuberculosis patients in the Azeri population of Iran. The TNF-308G/A single nucleotide polymorphism in the promoter region was genotyped by using the allele-specific PCR method in 200 healthy controls and 124 tuberculosis patients. The distribution of allele frequencies for TNF-? -308G/A polymorphism between control and tuberculosis patient groups was not significant (P-value = 0.058, OR = 1.5). Furthermore, no statistically significant association was found between TNF-? -308G/A genotype and resistance/susceptibility to TB (P-value = 0.102). Our results suggest that TNF-? -308G/A polymorphism has no measurable effect on the development of tuberculosis in Azeri population of Iran.

scholarly journals Analysis of Clinically Relevant Single-Nucleotide Polymorphisms by Use of Microelectronic Array Technology

2002 ◽  
Vol 48 (12) ◽  
pp. 2124-2130 ◽  
Author(s):  
Rosa Santacroce ◽  
Antonia Ratti ◽  
Francesco Caroli ◽  
Barbara Foglieni ◽  
Alessandro Ferraris ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphisms ◽  
Factor Vii ◽  
Restriction Enzyme Digestion ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Ret Protooncogene ◽  
Mismatched Dna ◽  
Specific Pcr ◽  
Pcr Products ◽  
Allele Specific

Abstract Background: Microelectronic DNA chip devices represent an emerging technology for genotyping. We developed methods for detection of single-nucleotide polymorphisms (SNPs) in clinically relevant genes. Methods: Primer pairs, with one containing a 5′-biotin group, were used to PCR-amplify the region encompassing the SNP to be interrogated. After denaturation, the biotinylated strand was electronically targeted to discrete sites on streptavidin-coated gel pads surfaces by use of a Nanogen Molecular Workstation. Allele-specific dye-labeled oligonucleotide reporters were used for detection of wild-type and variant sequences. Methods were developed for SNPs in genes, including factor VII, β-globin, and the RET protooncogene. We genotyped 331 samples for five DNA variations in the factor VII gene, &gt;600 samples from patients with β-thalassemia, and 15 samples for mutations within the RET protooncogene. All samples were previously typed by various methods, including DNA sequence analysis, allele-specific PCR, and/or restriction enzyme digestion of PCR products. Results: Analysis of amplified DNA required 4–6 h. After mismatched DNA was removed, signal-to-noise ratios were &gt;5. More than 940 samples were typed with the microelectronic array platform, and results were totally concordant with results obtained previously by other genotyping methods. Conclusions: The described protocols detect SNPs of clinical interest with results comparable to those of other genotyping methods.

scholarly journals Presence of the HPPD Inhibitor Sensitive 1 Gene and ALSS653N Mutation in Weedy Oryza sativa Sensitive to Benzobicyclon

Plants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1576
Author(s):  
Chad Brabham ◽  
Jason K. Norsworthy ◽  
Fidel González-Torralva
Keyword(s):  
Weedy Rice ◽  
Japonica Rice ◽  
Nucleotide Polymorphisms ◽  
Single Nucleotide ◽  
Herbicide Resistant ◽  
Rice Plants ◽  
Specific Pcr ◽  
Kasp Assay ◽  
Allele Specific ◽  
Allele Specific Pcr

Benzobicyclon has shown varying results in controlling weedy rice, including those with imidazolinone (IMI) resistance. Tolerance to benzobicyclon in cultivated japonica rice, but not indica or aus-like cultivars, is conferred by a fully functional HPPD Inhibitor Sensitive 1 (HIS1) gene. Herein, a diagnostic Kompetitive Allele Specific PCR (KASP) assay was developed to predict the HIS1 genotype of weedy rice plants from 37 accessions and correlated to their response to benzobicyclon in the field. Two-thirds of the 693 weedy rice plants screened were tolerant to benzobicyclon (371 g ai ha−1, SC formulation) at 30 days after treatment (DAT). Thirty-four percent of plants were homozygous for the HIS1 allele and 98% of these plants exhibited field tolerance. However, the his1 genotype did not always correlate with field data. Only 52% of his1 plants were considered sensitive, indicating that the single nucleotide polymorphisms (SNPs) chosen in the KASP assay are not a reliable tool in predicting his1 homozygous plants. In an additional experiment, 86% of the 344 plants with at least one copy of the ALSS653N trait harbored a HIS1 allele, suggesting fields infested with IMI herbicide-resistant weedy rice are unlikely to be controlled with benzobicyclon.

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