scholarly journals The frequency of terminal deletions sY160 among men with microdeletions of AZFc region Y-chromosome

1970 ◽  
Vol 21 ◽  
pp. 301-305
Author(s):  
I. Ye. Haiboniuk ◽  
M. Ya. Tyrkus ◽  
H. V. Makukh
Keyword(s):  
Y Chromosome ◽  
Molecular Genetic ◽  
Terminal Deletion ◽  
Molecular Genetic Study ◽  
Salting Out ◽  
Azfc Region ◽  
Specific Pcr ◽  
Allele Specific ◽  
Keywords Male Infertility ◽  
Azf Region

Aim. Determine the frequency of absence marker terminal deletions sY160 among men with microdeletions of AZFc region Y-chromosome. Methods. DNA from probands blood samples was isolated using a modified salting out method. Microdeletions of Y chromosome AZF region were analyzed using two multiplex PCR. The molecular-genetic study of terminal deletions (absence of sY160) was carried out using allele-specific PCR and analysed by electrophoresis in a 2 % agarose gel. Results. Among infertile men (1500 individuals), Y chromosome microdeletions were detected in 7% males: microdeletions of AZFa subregion in 1 %, AZFb subregion – 3 %, AZF(b+c) subregions – 15 %, AZFc subregion – 67 %. The presence of heterochromatin marker sY160 was confirmed in 39 cases (84.8 %) and absence of sY160 in 7 men (15.2 %). Absence of sY160 was detected in 2 men with AZFc microdeletion and in 5 men with AZFb+AZFc microdeletions. It is important to point out that terminal AZFc deletion was confirmed in 83.3 % of cases of AZFb+c microdeletions and only in 5.1 % of isolated AZFc microdeletions. Conclusions. Thus, among 15.2 % man with different AZF microdeletions of Y-chromosome the heterochromatin marker of terminal deletion sY160 was absents. The implementation of testing of marker of terminal deletion – sY160 may help to determine if the deletion corresponds to the b2/b4 pattern and to avoid biopsy in man which most likely not benefit from the surgical procedure. Keywords: male infertility, spermatogenesis, Y chromosome, AZF region, terminal deletions.

scholarly journals PCR đặc hiệu allele tích hợp công nghệ amplification refractory mutation system (Dsa-Pcr-Arms) phát hiện đột biến jak2 v617f và Carl trên bệnh nhân có hội chứng tăng sản tủy

2020 ◽  
Author(s):  
Tất Trung Ngô
Keyword(s):  
Molecular Genetic ◽  
Myeloproliferative Neoplasm ◽  
Jak2 V617f ◽  
Positive Sample ◽  
Amplification Refractory Mutation System ◽  
Study Cohort ◽  
Specific Pcr ◽  
Mutation System ◽  
Allele Specific ◽  
Allele Specific Pcr

BIDIRECTIONAL ALLELE SPECIFIC PCR INTEGRATED WITH AMPLIFICATION REFRACTORY MUTATION SYSTEM –(DSA-PCR-ARMS) FOR ANALYZING CARL AND JAK2 V617F MUTATIONS IN PATIENTS WITH MYELOPROLIFERATIVE NEOPLASM Bachground: Jak2 V617F and CARLmutation arerecognized as essential molecular genetic markers in diagnostic definition of myeloid proliferative neoplasm (MPN). Objective: to establish an income relevant Allele-specific-PCR-ARMS (DSA-PCR-ARMS) assays to identify Jak2 V617F, CARL mutations in MPN patients. Subjects: 64 MPN patients’ samples, one V617F positive referrance samples and one CALR mutant positive sample. Methods: dilution series of 100%, 50%, 25%, 5%, 0,5% and 0% mutant alleles were created as pseudo-samples establish the detection limits and the technical specificity of DSA-PCR-ARMS assays in identifying the mentioned genetics markers. Result: Our approach can detect presence of 0.5 % V617F mutant DNA fragment; beside that just by one gel-based PCR reaction, we can differentiate the deletion/insertion CARL carriers from the wild type counterpart. By applying our approachesto a study cohort of 50 MPN cases, a sensitivity of 64% and specificity of 100% was recorded for our Jak2 V617F detection assay and we also detected 3 CARL mutation case amongst the 10 V617F negative cases. Conclusion: The in-house of Jak2 V617F/CARL mutation detection protocols have been successfully established in our hospital. Key words: Jak2 V617F, CARL, myeloid proliferative neoplasm (MPN).

scholarly journals Analysis of fertility potential in men with severe azoospermia and oligospermia of various etiology

2018 ◽  
Vol 19 (3) ◽  
pp. 60-69
Author(s):  
T. A. Yamandi ◽  
L. V. Akulenko ◽  
N. Yu. Safina ◽  
I. I. Vityazeva ◽  
S. V. Bogolubov ◽  
...  
Keyword(s):  
Y Chromosome ◽  
Molecular Genetic ◽  
Normal Karyotype ◽  
Severe Oligozoospermia ◽  
Study Objective ◽  
Negative Results ◽  
Y Chromosome Microdeletions ◽  
Genetic Examination ◽  
Fertility Potential ◽  
Azf Region

The study objective is to evaluate the significance of the Y chromosome microdeletions for prediction of spermatozoa retrieval during testicle biopsy in men with severe azoospermia and oligozoospermia.Materials and methods. In total, 109 men aged 21 to 56 years (mean age 32.7 ± 0.2 years) with infertility in marriage were examined. Cytogenetic, special andrological, spermiological, and molecular genetic examinations were performed to evaluate non-genetic and genetic causes of infertility.Results. Normal karyotype and absence of AZF deletions were found in 75 men, presence of deletions – in 34. The frequencies of pathozoospermia forms were comparable in these groups. Spermatozoa were retrieved during biopsy in 47 (62.7 %) patients without Y chromosome microdeletions and only in 7 (20.6 %) patients with Y chromosome microdeletions. The men with AZF deletions were divided into 2 subgroups: men with complete AZF region deletions (n = 25) and men with partial AZF deletions (n = 9). Among men with complete deletions, azoospermia was diagnosed in 25 (100 %), spermatozoa were retrieved during biopsy in 2 (8 %); among men with partial deletions, azoospermia was diagnosed in 7 (77.8 %), severe oligozoospermia in 2 (22.2 %), spermatozoa were retrieved during biopsy in 5 (56 %). Then the patients were divided according to another criterion: 54 patients from whom spermatozoa were retrieved during biopsy and 55 men with negative results. Among patients with successful result of biopsy, Y chromosome microdeletions were identified in 7 (13 %); among patient with negative biopsy result – in 27 (49 %) (р < 0.01).Conclusion. Success rate of spermatozoa retrieval during testicle biopsy is significantly higher in men without AZF deletions (р < 0.01) than in men with deletions. Molecular genetic examination of Y chromosome microdeletions is recommended for men with azoospermia and severe oligozoospermia because it allows diagnosing of cause male infertility and predicting.

scholarly journals A High-Density Genetic Map Enables Genome Synteny and QTL Mapping of Vegetative Growth and Leaf Traits in Gardenia

2022 ◽  
Vol 12 ◽  
Author(s):  
Yang Cui ◽  
Baolian Fan ◽  
Xu Xu ◽  
Shasha Sheng ◽  
Yuhui Xu ◽  
...  
Keyword(s):  
Genetic Map ◽  
Molecular Genetic ◽  
Genetic Research ◽  
Genotyping By Sequencing ◽  
Leaf Traits ◽  
Coffea Canephora ◽  
High Density ◽  
Specific Pcr ◽  
True Hybrid ◽  
Allele Specific

The gardenia is a traditional medicinal horticultural plant in China, but its molecular genetic research has been largely hysteretic. Here, we constructed an F1 population with 200 true hybrid individuals. Using the genotyping-by-sequencing method, a high-density sex-average genetic map was generated that contained 4,249 SNPs with a total length of 1956.28 cM and an average genetic distance of 0.46 cM. We developed 17 SNP-based Kompetitive Allele-Specific PCR markers and found that 15 SNPs were successfully genotyped, of which 13 single-nucleotide polymorphism genotypings of 96 F1 individuals showed genotypes consistent with GBS-mined genotypes. A genomic collinearity analysis between gardenia and the Rubiaceae species Coffea arabica, Coffea canephora and Ophiorrhiza pumila showed the relativity strong conservation of LG11 with NC_039,919.1, HG974438.1 and Bliw01000011.1, respectively. Lastly, a quantitative trait loci analysis at three phenotyping time points (2019, 2020, and 2021) yielded 18 QTLs for growth-related traits and 31 QTLs for leaf-related traits, of which qBSBN7-1, qCD8 and qLNP2-1 could be repeatably detected. Five QTL regions (qCD8 and qSBD8, qBSBN7 and qSI7, qCD4-1 and qLLLS4, qLNP10 and qSLWS10-2, qSBD10 and qLLLS10) with potential pleiotropic effects were also observed. This study provides novel insight into molecular genetic research and could be helpful for further gene cloning and marker-assisted selection for early growth and development traits in the gardenia.

Y-chromosome variation in Indian native cattle breeds and crossbred population

2017 ◽  
Author(s):  
Indrajit Ganguly ◽  
Suchit Kumar ◽  
Sanjeev Singh ◽  
Monika Sodhi ◽  
Mukesh Bhakat
Keyword(s):  
Y Chromosome ◽  
Bos Taurus ◽  
Bos Indicus ◽  
Cost Effective ◽  
Cattle Breeds ◽  
Bull Semen ◽  
Specific Pcr ◽  
Allele Specific ◽  
Native Cattle ◽  
Male Specific

The paternally inherited Y chromosome markers have been used widely in population genetic studies to trace paternal lineages, to understand differences in migration pattern and populations admixture in animals. In the absence of crossing over, Y-chromosomal markers in the non-recombining male-specific region (MSY) are mostly transmitted as a haplotype. Recent studies of five polymorphic sites on DDX3Y, UTY and ZFY genes of bull MSY assisted in the identification of three haplotypes (Y1, Y2 and Y3) in contemporary cattle. Here we report the screening of five SNPs (ZFY9- 120> C/T; ZFY10- 655> C/T; DDX3Y1- 425>C/T; DDX3Y7 -123>C/T and UTY19-423>C/A) of bull MSY employing optimized and validated allele-specific PCR (AS-PCR) protocols that are useful in effective differentiation of bull/semen samples of Bos indicus and Bos taurus origin. Three haplotypes (Y1, Y2 and Y3) were identified in the present study by the screening of 181 bulls from 10 native cattle breeds and 50 HF crossbred. Y1 and Y2 haplotypes were restricted to HF crossbred with a frequency of 0.98 and 0.02, respectively. The high frequency of Y1 haplotype is possibly due to the occurrence of Y1 lineage predominantly in HF bulls. All the native cattle breeds were observed to have pure indicine haplotype (Y3). These cost effective AS-PCR protocols may be useful for reliable and accurate genotyping of Y-SNPs in diverse native cattle breeds, exotic and crossbred cattle populations.

scholarly journals COMPARISON OF DIFFERENT METHODS OF MOLECULAR-GENETIC ANALYSIS OF SOMATIC MUTATIONS IN K-RAS GENE IN PATIENTS WITH COLORECTAL CANCER

2012 ◽  
Vol 67 (2) ◽  
pp. 35-41 ◽  
Author(s):  
F. A. Amosenko ◽  
I. V. Karpov ◽  
A. V. Polyakov ◽  
S. P. Kovalenko ◽  
V. A. Shamanin ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
False Negative ◽  
Point Mutations ◽  
Real Life ◽  
Molecular Genetic Analysis ◽  
Ras Gene ◽  
Specific Pcr ◽  
Specificity And Sensitivity ◽  
Allele Specific ◽  
Allele Specific Pcr

Two approaches to somatic point mutations in 12 and 13 codones of K-ras gene were analyzed: PCR/SSCP/AСRS/sequencing and allele-specific PCR in the real-life regimen (Russian set «KRAS-7M»). The comparison was carried out on 62 examples of genomic DNA extracted from frozen colon carcinomas, which underwent manual dissection. The results obtained in two attempts were consistent in 95,2% (N=59). Specificity and sensitivity of K-ras mutations detection using «KRAS-7M» set were 100 and 96,4% respectively, and 94,1 and 100% respectievly using PCR/SSCP/AСRS/ automatic sequencing. False positive results were absent when detecting with «KRAS-7M» and accounted for 2 cases (5,9%) when using PCR/SSCP/ AСRS/automatic sequencing. The only false negative response (3,6%) was obtained analyzing mutations using «KRAS-7M».

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.

Challenges in the differential diagnosis of multiple endocrine neoplasia syndrome type 1 with isolated family hyperparathyroidism

2020 ◽  
Vol 98 (3) ◽  
pp. 218-225
Author(s):  
J. A. Krupinova ◽  
N. G. Mokrysheva ◽  
N. Y. Kalinchenko ◽  
A. K. Eremkina ◽  
A. N. Polyakov ◽  
...  
Keyword(s):  
Multiple Endocrine Neoplasia ◽  
Molecular Genetic ◽  
Pancreatic Neuroendocrine Tumor ◽  
Family Type ◽  
Dynamic Monitoring ◽  
Heterozygous Mutation ◽  
Molecular Genetic Study ◽  
Men 1 ◽  
Endocrine Neoplasia

Multiple endocrine neoplasia type 1 (MEN-1) is the most common cause of the hereditary type of primary hyperparathyroidism (PHPT). If a family type of PHPT is suspected, a dynamic monitoring of patients and their close relatives should be carried out throughout their lives. We present a clinical case of a family in which four members of a pedigree were diagnosed with familial isolated hyperparathyroidism (FIHP). The diagnosis was changed to MEN-1, because it appeared that one of the patients had pancreatic neuroendocrine tumor. Molecular genetic study of MEN1 by direct by means of Sanger sequencing revealed that six family members had a new heterozygous mutation in exon 9: s. 1252 G> T p. D418Y.

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