scholarly journals Polymorphism of Myostatin Gene in Intron 1 and 2 and Exon 3, and Their Associations with Yearling Weight, Using PCR-RFLP and PCR-SSCP Techniques in Zel Sheep

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Elena Dehnavi ◽  
Mojtaba Ahani Azari ◽  
Saeed Hasani ◽  
Mohammad Reza Nassiry ◽  
Mokhtar Mohajer ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Salting Out ◽  
Myostatin Gene ◽  
Blood Samples ◽  
Intron 1 ◽  
Pcr Rflp ◽  
Hardy Weinberg Equilibrium ◽  
Polymerase Chain ◽  
Intron 2

The aim of present study was to investigate myostatin gene polymorphism and its association with yearling weight records in Zel sheep using PCR-RFLP and PCR-SSCP methods. Blood samples were collected from 200 Zel sheep, randomly, and DNA was extracted using modified salting out method. Polymerase chain reaction was carried out to amplify 337, 222, and 311 bp fragments, respectively, comprising a part of exon 3, intron 1, and intron 2 of myostatin gene. In addition, exon 3 was digested by HaeIII enzyme under RFLP method, and introns 1 and 2 were studied using SSCP. Under RFLP method, all samples showed mm genotype. Under SSCP method, intron 1 was also monomorph but intron 2 was polymorph (AA, AB, and BB). The allelic frequencies for A and B were 75.5 and 24.5%, respectively. This locus was not in Hardy-Weinberg equilibrium (P<0.05), and there was no significant effect of myostatin gene on yearling weights.

scholarly journals Molecular Species Identification of Candida from Blood Samples of Intensive Care Unit Patients by Polymerase Chain Reaction – Restricted Fragment Length Polymorphism

2012 ◽  
Vol 4 (01) ◽  
pp. 001-004 ◽  
Author(s):  
Ramraj Vijayakumar ◽  
Sidhartha Giri ◽  
Anupma Jyoti Kindo
Keyword(s):  
Intensive Care Unit ◽  
Polymerase Chain Reaction ◽  
Intensive Care ◽  
Length Polymorphism ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Candida Spp ◽  
Blood Samples ◽  
Pcr Rflp ◽  
Polymerase Chain

ABSTRACT Introduction: Candida spp is an emerging cause of blood stream infections worldwide. Delay in speciation of Candida isolates by conventional methods and resistance to antifungal drugs (especially fluconazole, amphotericin B, etc.) in various Candida species are some of the factors responsible for the increase in morbidity and mortality due to candidemia. So, the rapid detection and identification of Candida isolates from blood is very important for the proper management of patients having candidemia. Materials and Methods: In this study, we have used polymerase chain reaction (PCR) - restriction fragment length polymorphism (RFLP) as a method for the speciation of Candida isolates from blood samples of intensive care unit (ICU) patients. PCR was used to amplify the ITS-1 and ITS-2 regions of Candida spp using universal primers ITS-1 and ITS-4. The amplified product was digested using Msp I restriction enzyme by RFLP. Results and Discussion: The method PCR-RFLP helped in identifying five medically important Candida spp (C. tropicalis, C. albicans, C. parapsilosis, C. krusei and C. glabrata) from blood. This method is rapid, reliable, easy and cost-effective and can be used in routine laboratory diagnostics for the rapid identification of Candida isolates from blood. Conclusion: PCR-RFLP is an easy, rapid and highly valuable tool which can be used in routine diagnostic laboratories to speciate Candida isolates obtained from blood. This rapid method of speciation will help clinicians to decide on empirical therapy in candidemia cases before antifungal susceptibility results are available.

scholarly journals Polymorphisms in CCR5Δ32 and Risk of HIV-1 Infection in the Southeast of Caspian Sea, Iran

2017 ◽  
Vol 2017 ◽  
pp. 1-5 ◽  
Author(s):  
Zahra Heydarifard ◽  
Alijan Tabarraei ◽  
Abdolvahab Moradi
Keyword(s):  
Polymerase Chain Reaction ◽  
Hiv Infection ◽  
Caspian Sea ◽  
Chain Reaction ◽  
Blood Samples ◽  
Infected People ◽  
Significant Difference ◽  
Hardy Weinberg Equilibrium ◽  
Polymerase Chain ◽  
Hiv 1

Prevalence of CCR5Δ32 among blood samples of more than 400 healthy and HIV-1-infected people was investigated in Iran. Polymerase chain reaction (PCR) following DNA extraction was used. Desired frequency was analyzed by Hardy–Weinberg equilibrium (HWE) analysis and SPSS 16.0 software to harvest the results. The prevalence of CCRΔ32 heterozygote genotype was 3% in healthy people and 0.7% in HIV-1–infected individuals. There was no homozygote CCR5Δ32 in both groups, and the allele Δ32 was only observed in 1.5% and 0.36% of healthy and HIV-1–infected participants, respectively. Therefore according to this study, the frequency of the allele CCR5Δ32 indicates no significant difference between either groups (p=0.18) and it sounds that the mentioned mutation in heterozygote people would not affect their susceptibility against HIV infection. Genotyping trial in Iranians with HIV infection is supposed to be helpful as a matter of prognostic purposes.

scholarly journals Molecular surveillance of Theileria ovis, Theileria lestoquardi and Theileria annulata infection in sheep and ixodid ticks in Iran

2013 ◽  
Vol 80 (1) ◽  
Author(s):  
Gholamreza Razmi ◽  
Saeed Yaghfoori
Keyword(s):  
Polymerase Chain Reaction ◽  
Salivary Glands ◽  
Nested Pcr ◽  
Ixodid Ticks ◽  
Chain Reaction ◽  
Blood Samples ◽  
Theileria Lestoquardi ◽  
Blood Smears ◽  
Pcr Rflp ◽  
Polymerase Chain

A molecular study was undertaken to detect Theileria ovis, Theileria lestoquardi and Theileria annulatain sheep and tick vectors. Investigation was conducted from 2010 to 2011 in the south of Khorasan Razavi Province, Iran. A total of 150 blood samples were collected from 30 different sheep flocks. In addition, ixodid ticks were sampled from the same flocks. The stained blood smears were microscopically examined for the presence of piroplasms and a semi-nested polymerase chain reaction-restriction (PCR) was used for subsequent molecular speciation. Salivary glands were isolated from the ticks and subsequently analysed by semi-nested PCR. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to differentiate between T. lestoquardi and T. annulata from PCR-positive samples. Theileria species infection was microscopically detected in 18.6% of blood smears. The presence of T. ovis and T. lestoquardi or T. annulata was detected by semi-nested PCR in 58.6% and 6.6% of blood samples respectively. In total, 169 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 155; 91.7% of the total), followed by Hyalomma anatolicum anatolicum (n = 8; 4.7%) and Hyalomma marginatum turanicum (n = 6; 3.5%). From an organ pooling of 33 ticks, three pools of salivary glands from R. turanicus were positive for Theileria species by semi-nested PCR. Of the three R. turanicus samples testing positive for Theileria species, two (6.1%) were positive for T. ovis and one (3.0%) for T. lestoquardi or T. annulata. Amongst the 11 PCR-positive samples for T. lestoquardi or T. annulata, 10 were positive for T. lestoquardi and one sample was positive for both T. lestoquardi and T. annulata using PCR-RFLP. The results also demonstrated that PCR-RFLP could be used for the detection of T. ovis. Based on the results, it can be concluded that T. ovis has a higher prevalence than T. lestoquardi, and that R. turanicus could be a possible vector for T. ovis and T. lestoquardi. Finally, the PCR-RFLP based on Msp1 restriction enzyme is a simple method for differentiation of Theileria species in sheep and ixodid ticks.

scholarly journals Polimorfismo no gene GH1-PstI associado a características corporais de linhagens de tilápia-do-nilo

2009 ◽  
Vol 44 (6) ◽  
pp. 599-604 ◽  
Author(s):  
Danielly Veloso Blanck ◽  
Eliane Gasparino ◽  
Ricardo Pereira Ribeiro ◽  
Débora Sommer Marques
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment ◽  
Oreochromis Niloticus ◽  
Chain Reaction ◽  
Intron 1 ◽  
Pcr Rflp ◽  
Polymerase Chain

O objetivo deste trabalho foi determinar a associação de polimorfismos no gene do hormônio crescimento GH1 às características corporais, em linhagens de tilápia-do-nilo (Oreochromis niloticus). Foram coletados fragmentos da nadadeira caudal de exemplares das linhagens, aos cinco meses de idade, para as análises de "polymerase chain reaction-restriction fragment lenght polymorphism" (PCR-RFLP). Foram realizadas as seguintes mensurações: comprimento total, comprimento padrão, altura, largura e comprimento da cabeça. Realizou-se a amplificação de um fragmento com 652 pb do gene GH1, com subsequente restrição com a enzima PstI. Para a análise de associação do marcador molecular com as características quantitativas, utilizou-se o procedimento GLM do SAS. O polimorfismo descrito para o íntron 1, do gene GH1 da tilápia-do-nilo, apresentou correlação significativa com o comprimento total, comprimento padrão, altura e largura corporal. Foi verificado que o genótipo PstI+/- está associado ao melhor crescimento, independentemente da linhagem. A associação verificada pode ter ocorrido em razão do efeito direto da regulação do próprio gene GH.

Investigation of Mutations in Exon 12 Of MYH7, 16 Of β-MYBPC3 and 2 Of TCAP Gene in Dogs with Dilated Cardiomyopathy using PCR-SSCP Technique

2021 ◽  
Author(s):  
R.B. Vishnurahav ◽  
S. Ajithkumar ◽  
Usha Narayana Pillai ◽  
N. Madhvan Unny ◽  
K.D. John Martin ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dilated Cardiomyopathy ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Multifunctional Protein ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Intron 1 ◽  
Cardiac Disorders ◽  
Polymerase Chain

Background: Dilated cardiomyopathy is the important myocardial disease and one of the most common cause of death in the medium to large size dog breeds worldwide. The disease is characterized by dilatation of cardiac chambers and thinning of walls leads to systolic failure. Mutations in some sarcomere genes leads to cardiomyopathy in humans. Sarcomere is an important multifunctional protein network involved in the signal reception and transduction. Mutations in β-MYH7, MYBPC3 and TCAP genes produce alterations in the morphology of heart (hypertrophy or dilatation).Methods: In this study twenty apparently healthy and twenty five dogs with dilated cardiomyopathy (DCM) were selected from patients reported or referred to University Veterinary Hospital and Teaching Veterinary Clinical Complex, Mannuthy (2015-2017) based on the clinical examination, radiographic, electrocardiographic, haematobiochemical and echocardiographic studies cardiac disorders (Dilated cardiomyopathy and hypertrophic cardiomyopathy) were confirmed.Result: In the present study we investigated genetic alterations of exon 12 of MYH7, 16 of β-MYBPC3 and 2 of TCAP gene in dogs by polymerase chain reaction -single stranded confirmation of polymorphism (PCR-SSCP). Polymerase chain reactions were analysed using acrylamide gel and samples with different pattern of bands were sequenced. Polymerase chain reaction-SSCP showed different migration of band pattern in the intron 1 of TCAP gene in one sample.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

scholarly journals Extração de DNA a partir de sangue humano coagulado para aplicação nas técnicas de genotipagem de antígenos leucocitários humanos e de receptores semelhantes à imunoglobulina

2009 ◽  
Vol 42 (6) ◽  
pp. 651-656 ◽  
Author(s):  
Daniela Maira Cardozo ◽  
Gláucia Andréia Guelsin ◽  
Samaia Laface Clementino ◽  
Fabiano Cavalcante de Melo ◽  
Marco Antônio Braga ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Killer Cell ◽  
Human Leukocyte ◽  
Reaction Sequence ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Salting Out ◽  
Leukocyte Antigens ◽  
Polymerase Chain ◽  
Made In

O objetivo deste estudo foi padronizar uma metodologia de extração de DNA de alta qualidade a partir de amostras de sangue coagulado. Quarenta e oito amostras de sangue humano coagulado foram utilizadas para a extração de DNA pelo kit comercial EZ-DNA® (Biological Industries, Beit Haemek, Israel), pelo kit de coluna Neoscience® (One Lambda Inc., San Diego, CA) e pelo método modificado de salting out. Apenas o método de salting out foi capaz de extrair altas concentrações de DNA (média, 180ng/µL), as quais foram medidas pelo detector de fluorescência Qubit® (Invitrogen, USA). Este método permitiu a amplificação dos genes HLA (human leukocyte antigens) pela tecnologia PCR-SSO (polymerase chain reaction - specific sequence of oligonucleotides) Luminex, a qual exige DNA de boa qualidade, e de genes KIR (killer cell immunoglobulin-like receptors) pela técnica made in house PCR-SSP (polymerase chain reaction-sequence specific of primers), a qual demanda uma concentração específica de DNA (10ng/µL). Concluímos que a técnica de salting out modificada foi muito eficiente, simples e rápida para a extração de DNA de amostras de sangue humano coagulado, com o objetivo de realizar a genotipagem de genes HLA e KIR.

scholarly journals Identifikasi Keragaman Gen DGAT1 serta Asosiasinya terhadap Karakteristik Karkas dan Sifat Perlemakan Domba

2019 ◽  
Vol 6 (2) ◽  
pp. 259
Author(s):  
Asep Gunawan ◽  
Ratna Sholatia Harahap ◽  
Kasita Listyarini ◽  
Cece Sumantri
Keyword(s):  
Polymerase Chain Reaction ◽  
Single Nucleotide Polymorphism ◽  
Fragment Length Polymorphism ◽  
Nucleotide Polymorphism ◽  
Chain Reaction ◽  
Single Nucleotide ◽  
Dgat1 Gene ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

ABSTRAK Karakteristik karkas dan sifat perlemakan pada daging domba dikontrol oleh banyak gen salah satunya gen DGAT1 (Diacylglycerol Acyltransferasel 1). Penelitian ini bertujuan mengidentifikasi SNP (Single Nucleotide Polymorphism) gen DGAT1 pada titik mutasi g.8539 C>T dan asosiasinya terhadap karakteristik karkas dan sifat perlemakan pada domba Indonesia. Total sampel domba yang digunakan sebanyak 150 buah terdiri dari 35 sampel domba compass agrinak (DCA), 36 sampel domba barbados cross (DBC), 41 sampel domba komposit garut (DKG), 20 sampel domba ekor gemuk (DEG), dan 18 sampel domba ekor tipis (DET). Karakteristik karkas dan sifat perlemakan diukur dari domba jantan berumur 10-12 bulan. Identifikasi keragaman DGAT1|ALuI dianalisis dengan metode PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Hasil keragaman gen DGAT1 bersifat polimorfik dalam DET dan DEG, sedangkan DCA, DBC, dan DKG bersifat monomorfik. Dua genotipe disebut CC dan  CT ditemukan dalam DET dan DEG. Titik mutasi gen DGAT1 berasosiasi (P<0.05) dengan karakteristik karkas, yaitu bobot dan panjang karkas. Selain itu, keragaman gen DGAT1 juga berasosiasi signifikan (P<0.05) dengan asam lemak jenuh, yaitu asam stearat (C18:0) dan asam arakidat (C20:0) dan asam lemak tak jenuh tunggal, yaitu asam oleat (C18:1n9c). Gen DGAT1 memiliki kontribusi dalam karakteristik karkas dan komposisi asam lemak pada domba.Kata Kunci: domba, gen DGAT1, karakteristik karkas, PCR-RFLP, sifat perlemakan                                                              ABSTRACT            Characteristic of carcass and fatness traits of sheep is regulated by many genes such as DGAT1 (Diacylglycerol Acyltransferasel 1) gene. The research was aimed to investigate SNP (Single Nucleotide Polymorphism) of DGAT1 and its association with characteristic of carcass and fatness traits in Indonesian sheep. A total sample of sheeps used 150 rams of 10–12 months consisted 35 samples of compas agrinak sheep (CAS), 36 of barbados cross (BCS), 41 of garut composite (GCS), 20  of javanese fat tailed (JFT), and 18 of javanese thin tailed (JTT). Identification variant of DGAT1|ALuI were performed by PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). The results of polymorphism of DGAT1 were found in JTT and JFT. However, SNP of DGAT1 in CAS, BCS and GCS were monomorfic. Two genotype namely CC and CT were found in JTT and JFT populations. A SNP of the DGAT1 was associated (P<0.05) with characteristic of carcass, including weight and length of carcass. The variant of DGAT1 was associated too with saturated fatty acids (SFA) including stearic acid (C18:0) and arachidic acid (C20:0), and mono unsaturated fatty acid (MUFA) including oleic acid (C18:1n9c). The DGAT1 gene was contribute to characteristic carcass and fatty acid composition in sheep.Keywords: DGAT1 gene, characteristic carcass, fatness traits, PCR-RFLP, sheep

scholarly journals Etude de l’implication du polymorphisme fonctionnel Arg72Pro du gène TP53 dans la survenue du cancer colorectal et du carcinome Basocellulaire dans la population de l’Ouest Algérien

2017 ◽  
Vol 1 (2) ◽  
Author(s):  
Rym Abderrahmane ◽  
Lotfi Louhibi ◽  
Amina Boubekeur ◽  
Fatima Zohra Moghtit ◽  
Abdellah Boudjema ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment ◽  
Cancer Colorectal ◽  
Facteurs De Risque ◽  
Chain Reaction ◽  
Biologie Moléculaire ◽  
Pcr Rflp ◽  
Cycle Cellulaire ◽  
Gene Tp53 ◽  
Polymerase Chain

Introduction - Le gène TP53 a fait l’objet de très nombreux travaux. Sa nature polymorphe et son rôle central dans la régulation du cycle cellulaire ont mis en évidence son potentiel de gène candidat dans la susceptibilité et la survenue de différents cancers. Bien que plusieurs polymorphismes du gène TP53 aient été étudiés comme facteurs de risque pour différents cancers, le plus largement étudié est le polymorphisme Arg72Pro (rs1042522), à l’origine d’une substitution d’une Arginie (Arg) en Proline (Pro) ou inversement, connu pour ses différentes fonctions biologiques.Notre objectif est d’explorer la participation du polymorphisme (SNP) Arg72Pro du gène TP53, dans l’expressivité du cancer colorectal (CCR) et du carcinome basocellulaire (CBC) dans une population de l’Ouest Algérien.Matériels et Méthodes - Le déséquilibre de transmission allélique de ce SNP a été réalisé sur une population constituée de 116 individus atteints de CCR, 50 sujets atteints de CBC et enfin une population contrôle constituée de 121 individus sains,par la technique de Biologie Moléculaire PCR/RFLP (Polymerase Chain Reaction/Restriction Fragment LengthPolymorphism).Résultats - L’association du marqueur Arg72Pro avec le CBC a montré une augmentation significative de l’allèle Pro chez les cas par rapport aux contrôles (54% vs 46%, OR=8,85 [4,98-16,03], p<10-7). Cet allèle semble conférer un risque important dedévelopper le CBC. Par ailleurs, les résultats n’ont montré aucune différence significative entre les sujets atteints de CCR et les contrôles, ce qui pourrait exclure son implication dans la survenue du CCR dans notre population (p>0.05).Conclusion - Le polymorphisme Arg72Pro du gène TP53 peut être considéré comme marqueur de risque pour le CBC mais pas pour le CCR.

scholarly journals Molecular detection and occurrence of equine theileriosis in Arabian horses in Al-Najaf province/Iraq

2020 ◽  
Vol 57 (3) ◽  
pp. e166996
Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Extraction ◽  
Molecular Detection ◽  
Molecular Technique ◽  
Chain Reaction ◽  
Blood Samples ◽  
Equine Piroplasmosis ◽  
Polymerase Chain

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.

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