scholarly journals Sanitary Status of the Grapevine Germplasm Collection in Republic of Srpska

2017 ◽  
Vol 17 (2) ◽  
pp. 143
Author(s):  
Duška Delić ◽  
Biljana Lolić ◽  
Gordana Đurić ◽  
Tatjana Jovanović-Cvetković
Keyword(s):  
Nested Pcr ◽  
Germplasm Collection ◽  
Elisa Test ◽  
Grapevine Fanleaf Virus ◽  
Pcr Rflp ◽  
Near Future ◽  
Infected Grapevine ◽  
Rflp Assay ◽  
Das Elisa

In July 2015, 179 grapevine plants belonging to 16 grapevine autochthonous cultivars were assessed for sanitary status using DAS ELISA test for the presence of: Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated virus 1 (GLRaV-1), Grapevine leafroll-associated virus 2 (GLRaV-2)and Grapevine leafroll-associated virus 3 (GLRaV-3). Furthermore, surveyfor the phytoplasma presence and laboratory analyses using nested-PCR/RFLP assay was conducted at the beginning of September 2015 on grapevine cultivars which were not positive in DAS ELISA test for the presence of the four viruses. Out of 179 tested plants with DAS ELISA test, 146 (81%) were positive for the presence of at least one virus. The most widespread viruses were GFLaV- 1 and GFLaV- 3 with approximately 80 % of grapevines infected. Nested–PCR/RFLP assay showed that out of 33 tested samples 2 were positive for the presence of phytoplasmas from 16SrXII group. Sanitation of infected grapevine cultivars is needed in near future.

scholarly journals Evaluation of sanitary status of grapevines in the Czech Republic

2011 ◽  
Vol 49 (No. 2) ◽  
pp. 63-66 ◽  
Author(s):  
P. Komínek ◽  
V. Holleinová
Keyword(s):  
Czech Republic ◽  
Grapevine Virus ◽  
Arabis Mosaic Virus ◽  
The Czech Republic ◽  
Grapevine Virus A ◽  
Grapevine Virus B ◽  
Near Future ◽  
Infected Grapevine ◽  
Das Elisa ◽  
Propagation Material

A survey was made to evaluate sanitary status of grapevines in the Czech Republic with regard to occurrence of economically important viruses. Propagation material of 109 grapevine clones was tested for presence of Grapevine fanleaf virus, Arabis mosaic virus, Grapevine leafroll-associated virus 1, Grapevine leafroll-associated virus 3, Grapevine virus A, Grapevine virus B and Grapevine fleck virus. Dormant canes were collected and cortical scrappings were analyzed by DAS-ELISA. All seven viruses tested were found to be widely spread in Czech propagation material of grapevine. From 330 individual vines tested, 148 vines were found to be infected with at least one virus. From 109 clones tested, in 98 clones at least one vine negative for tested pathogens was found. Such vines were promoted as candidate plants into screenhouse in Faculty of Horticulture Lednice and will be further tested by other methods. Sanitation of infected grapevine clones is needed in near future.

scholarly journals Grapevine Pinot gris virus infecting grapevines in Romania - Short Communicaiton

2021 ◽  
Vol 48 (No. 1) ◽  
pp. 47-50
Author(s):  
Ionela-Catalina Guta ◽  
Elena-Cocuta Buciumeanu
Keyword(s):  
Clonal Selection ◽  
Germplasm Collection ◽  
High Rate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Grapevine Fanleaf Virus ◽  
Mother Plants ◽  
Grapevine Pinot Gris Virus ◽  
New Varieties ◽  
Das Elisa ◽  
Propagation Material

Grapevine Pinot gris virus (GPGV) has been identified in many grape growing countries of the world since 2012. The aim of this work was to investigate the presence of GPGV on some accessions collected from a germplasm collection, in addition to the propagation material and clonal selection samples. During 2019–2020, a total of 199 samples have been analysed by a double antibody sandwich – enzyme-linked immunosorbent assay (DAS-ELISA) for the presence of GPGV, Grapevine fanleaf virus (GFLV), Grapevine leafroll-associated virus-1+3 (GLRaV-1+3) and Grapevine fleck virus (GFkV). Among them, 107 samples (53.76%) showed a GPGV-infection, associated with or without symptoms on the leaves (deformations, chlorosis, mosaic, wrinkles) or stunting plants. The distribution of infected varieties showed a high rate of infection in old varieties (37.38%), followed by clones (32.71%), rootstocks (11.21%), clonal selections (9.35%) and new varieties (9.35%). The tests revealed the association of GPGV with GFkV (5 cases) and GLRaV-1+3 (2 cases). GPGV should be included in the rules of grapevine certification schemes for the production of virus-free mother plants.

Sanitation of Autochthonous Grapevine Varieties from Algeria by Chemotherapy

2020 ◽  
Vol 55 (1) ◽  
pp. 43-50
Author(s):  
N. Aiter ◽  
A. Lehad ◽  
B. Haddad ◽  
A. Taibi ◽  
S. Meziani ◽  
...  
Keyword(s):  
Virus Infection ◽  
Germplasm Collection ◽  
Elisa Test ◽  
Shoot Tips ◽  
Breeding Programs ◽  
Significant Rate ◽  
Das Elisa

Several grapevine viruses were reported in Algeria and especially in grapevine germplasm collection, therefore it is a great challenge to free these varieties from virus infection before any breeding programs. Our study focused on the development of chemotherapy on autochthonous varieties collected in the grapevine germplasm collection of ITAFV. All these varieties were tested by DAS-ELISA and the presence of GLRaV-3 and GFLV was confirmed in all used samples for the sanitation. After 8 weeks of shoot tips in vitro culture in a modified M S medium containing ribavirin, DAS-ELISA test revealed that GLRaV-3 was completely eliminated and GFLV to a significant rate.

scholarly journals Human papillomavirus detection and typing using a nested-PCR-RFLP assay

2011 ◽  
Vol 15 (5) ◽  
pp. 467-472 ◽  
Author(s):  
Janaina Coser ◽  
Thaís da Rocha Boeira ◽  
André Salvador Kazantzi Fonseca ◽  
Nilo Ikuta ◽  
Vagner Ricardo Lunge
Keyword(s):  
Human Papillomavirus ◽  
Nested Pcr ◽  
Pcr Rflp ◽  
Rflp Assay

scholarly journals Human papillomavirus detection and typing using a nested-PCR-RFLP assay

2011 ◽  
Vol 15 (5) ◽  
pp. 467-472 ◽  
Author(s):  
Janaina Coser ◽  
Thaís da Rocha Boeira ◽  
André Salvador Kazantzi Fonseca ◽  
Nilo Ikuta ◽  
Vagner Ricardo Lunge
Keyword(s):  
Human Papillomavirus ◽  
Nested Pcr ◽  
Pcr Rflp ◽  
Rflp Assay

PCR-RFLP Detection od PAI-2 Gene Variants: Prevelence in Ethnic Groups and Disease Relationship in patients Undergoing corony Angiography

1997 ◽  
Vol 77 (05) ◽  
pp. 0955-0958 ◽  
Author(s):  
Carole A Foy ◽  
Peter J Grant
Keyword(s):  
Gene Variants ◽  
Genotype Distribution ◽  
Pima Indians ◽  
Significant Difference ◽  
Pcr Rflp ◽  
History Of ◽  
Caucasian Patients ◽  
Clinical Conditions ◽  
Rflp Assay ◽  
Coronary Atheroma

SummaryPAI-2 is a fibrinolytic inhibitor produced predominantly by monocytes. Most PAI-2 is intracellular making study in clinical conditions difficult. Abnormalities in production may be associated with inflammation and fibrinolysis at sites of tissue damage such as the atherosclerotic plaque.PAI-2 gene variants have been described: variant A consists of Asn120, Asn404 and Ser413 and variant B consists of Asp120, Lys404 and Cys413. We designed a PCR-RFLP assay using primers spanning the region containing Asn/Lys404 and Ser/Cys413. Variant B contains an Mwol restriction site. We analysed 302 Pima Indians and 286 healthy Caucasian volunteers. To investigate relationships between genotype and vascular disease we analysed 333 Caucasian patients undergoing coronary angiography.Gene variant B was more common in the Pimas than in Caucasians (p <0.0001). There was no significant difference in genotype distribution between the volunteers and patients. In the patients there was no association between genotype and either a history of MI or extent of coronary atheroma.

scholarly journals Molecular characteristics of HBV infection among blood donors tested HBsAg reactive in a single ELISA test in southern China

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xianlin Ye ◽  
Tong Li ◽  
Ran Li ◽  
Heng Liu ◽  
Junpeng Zhao ◽  
...  
Keyword(s):  
Viral Load ◽  
Hepatitis B ◽  
Nested Pcr ◽  
Blood Donors ◽  
Southern China ◽  
Core Promoter ◽  
Elisa Test ◽  
Hbv Infection ◽  
Molecular Characteristics ◽  
Blood Samples

Abstract Background Hepatitis B virus (HBV) infection is a major concern for blood safety in high-prevalence HBV countries such as China. In Shenzhen, dual hepatitis B surface antigen (HBsAg) enzyme-linked immunosorbent assays (ELISAs) have been adopted in parallel with nucleic acid testing (NAT) for donors for over a decade. A small proportion of blood donors test reactive (R) for HBsAg but negative through routine NAT, which can lead to HBV infection with an extremely low viral load. Objectives We aimed to investigate and analyze the molecular characteristics of HBV among blood donors that tested HBsAg R in a single ELISA test. Methods Blood donations were evaluated in this study if confirmed HBsAg R through one of two ELISA kits. Samples with non-reactive (NR) results by NAT were collected and tested for HBsAg by chemiluminescent microparticle immunoassay (CLIA) with a neutralization test. The level of HBsAg was further assessed by electrochemiluminescence immunoassay (ECLIA). The viral basic core promoter (BCP) and pre-core (PC) and S regions were amplified by nested PCR. Quantitative real-time PCR (qPCR) for viral load determination and individual donation (ID)-NAT were adopted simultaneously. HBsAg was confirmed with CLIA, ECLIA, nested PCR, qPCR, and ID-NAT. Results Of the 100,252 donations, 38 and 41 were identified as HBsAg R with Wantai and DiaSorin ELISA kits, respectively. Seventy-nine (0.077%, 79/100,252) blood samples with ELISA R-NR and NAT NR results were enrolled in the study. Of these, 17 (21.5%,17/79) were confirmed as HBsAg-positive. Of the 14 genotyped cases, 78.6% (11/14) were genotype B, and C and D were observed in two and one sample, respectively. Mutations were found in the S gene, including Y100C, Y103I, G145R, and L175S, which can affect the detection of HBsAg. A high-frequency mutation, T1719G (93.3%), was detected in the BCP/PC region, which reduced the viral replication. Conclusion A small number of blood samples with HBsAg ELISA R-NR and NAT NR results were confirmed as HBV infection, viral nucleic acids were found in most of the samples through routine NAT methods. It is necessary to employ more sensitive and specific assays for the detection of HBV infection among blood donors.

PCR‐RFLP assay confirms the existence of different mitochondrial lineages of Taenia hydatigena including a possible geographically restricted group

2021 ◽  
Author(s):  
John Asekhaen Ohiolei ◽  
Hong‐Bin Yan ◽  
Li Li ◽  
Mughees Aizaz Alvi ◽  
Rosline James Muku ◽  
...  
Keyword(s):  
Taenia Hydatigena ◽  
Pcr Rflp ◽  
Restricted Group ◽  
Rflp Assay

Sex identification PCR–RFLP assay tested in eight species of Sebastes rockfish

2020 ◽  
Vol 12 (4) ◽  
pp. 541-544
Author(s):  
Felix Vaux ◽  
Hannah M. Aycock ◽  
Sandra Bohn ◽  
Leif K. Rasmuson ◽  
Kathleen G. O’Malley
Keyword(s):  
Sex Identification ◽  
Pcr Rflp ◽  
Rflp Assay

scholarly journals PCR-based genotyping assays to detect germline APC variant associated with hereditary gastrointestinal polyposis in Jack Russell terriers

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kyoko Yoshizaki ◽  
Akihiro Hirata ◽  
Hiroyuki Matsushita ◽  
Naohito Nishii ◽  
Mifumi Kawabe ◽  
...  
Keyword(s):  
Mutant Allele ◽  
False Negative ◽  
Disease Onset ◽  
Pcr Amplification ◽  
Buccal Swab ◽  
Wild Type ◽  
Gastrointestinal Polyposis ◽  
Pcr Rflp ◽  
Formalin Fixed Paraffin Embedded ◽  
Rflp Assay

Abstract Background The prevalence of gastrointestinal (GI) neoplastic polyps in Jack Russell terriers (JRTs) has increased in Japan since the late 2000s. Recently, we demonstrated that JRTs with GI polyps harbor identical germline variant in the APC gene (c.[462_463delinsTT]) in the heterozygous state. Thus, this disease is an autosomal dominant hereditary disorder. Although the affected JRTs have distinct features, such as the development of multiple GI polyps and an early age of disease onset, genetic testing is indispensable for a definitive diagnosis. Here, polymerase chain reaction (PCR)-based assays capable of detecting germline APC variant were designed and validated using synthetic wild-type and mutant DNAs and genomic DNAs from carrier and non-carrier dogs. Result First, the PCR-restriction fragment length polymorphism (PCR-RFLP) assay was developed by taking advantage of the germline APC variant creating a new restriction site for MseI. In the PCR-RFLP assay, the 156-bp region containing the variant site was amplified by PCR and subsequently digested with MseI, yielding diagnostic 51 and 58 bp fragments from the mutant allele and allowing determination of the APC genotypes. It was possible to determine the genotypes using genomic DNA extracted from the peripheral blood, buccal swab, or formalin-fixed paraffin-embedded tissue. Next, a TaqMan duplex real-time PCR assay was developed, where a 78-bp region flanking the variant was amplified in the presence of wild-type allele- and mutant allele-specific fluorescent probes. Using blood-derived DNA, altogether 40 cycles of PCR amplification determined the APC genotypes of all examined samples by measuring the fluorescence intensities. Importantly, false-positive and false-negative errors were never detected in both assays. Conclusion In this study, we developed highly reliable genetic tests for hereditary GI polyposis in JRTs, providing accurate assessment of the presence of the causative germline APC variant. The genotyping assays could contribute to the diagnosis and prevention of hereditary GI polyposis in dogs.

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