Differentiation of infectious bronchitis virus vaccine strains Ma5 and 4/91 by TaqMan real-time PCR

2017 ◽  
Vol 20 (3) ◽  
pp. 599-601 ◽  
Author(s):  
T. Stenzel ◽  
D. Dziewulska ◽  
M. Śmiałek ◽  
B. Tykałowski ◽  
J. Kowalczyk ◽  
...  
Keyword(s):  
Real Time ◽  
Virus Vaccine ◽  
Infectious Bronchitis Virus ◽  
Real Time Pcr ◽  
Cross Reactivity ◽  
Specific Primers ◽  
Assay Sensitivity ◽  
Infectious Bronchitis ◽  
Molecular Assays ◽  
Rapid Molecular Assays

Abstract The aim of this study was to develop rapid molecular assays for differentiating vaccine strains Ma5 and 4/91 of the infectious bronchitis virus (IBV). Specific primers and probes for S1 and N genes were designed based on the nucleotide sequences of both vaccine strains. Cross-reactivity was not observed. Assay sensitivity was 2.373 × 103 copies of the Ma5 strain, and 3.852 x 103 copies of the 4/91 strain. Samples belonging to a known genotype demonstrated that the designed assays supported rapid and sensitive detection of Ma5 and 4/91 vaccine strains of IBV.

scholarly journals Developing, Modifying, and Validating a TaqMan Real-Time PCR Technique for Accurate Identification of Leishmania Parasites Causing Most Leishmaniasis in Iran

2021 ◽  
Vol 11 ◽  
Author(s):  
Reza Fotouhi-Ardakani ◽  
Seyedeh Maryam Ghafari ◽  
Paul Donald Ready ◽  
Parviz Parvizi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Leishmania Major ◽  
Taqman Probe ◽  
Amino Acid Permease ◽  
Specific Primers ◽  
Accurate Identification ◽  
Parasitic Load ◽  
Species Specific

Many laboratory methods are used to diagnose leishmaniasis because it is characterized by varied symptoms and caused by different Leishmania species. A quantitative real-time PCR method based on a TaqMan probe was developed and modified for accurate identification of human cutaneous leishmaniasis (caused by Leishmania major or Leishmania tropica) from endemic areas of Iran. Two gene regions of amino acid permease 3 (AAP3) and cytochrome oxidase II (COII) were considered. Six new sets of species-specific primers and probes were designed. A total of 123 samples were examined and employed to evaluate and validate real-time PCR. According to parasitic load of the genesig®Leishmania Advanced Standard Kit, a serial dilution of purified plasmid (2–2×107 copies/reaction) was prepared under the same conditions for both genes. Specific primers and probes were able to detect three and six parasite copies in AAP3 and COII genes, respectively, and were able to detect three copies of parasites for L. major and L. tropica. The sensitivities of the reference kit and our method were 98.7 and 98.1%, respectively, and specificity was 100% for detecting parasite genomes in all assays. Designed primers and probes performed well in terms of efficiency and regression coefficient. For AAP3 and COII genes, respectively, the linear log range was 7 and the correlation coefficient (R2) was 0.749 and 0.996 for the reference kit using the standard generated curve and 0.98 and 0.96 with serial dilutions of parasite DNA. This research detected L. major and L. tropica definitely and opens the horizon for the other scientists in the multiplex reactions in designing and optimization of the conditions in silico and in vivo.

scholarly journals Characterization of a novel live attenuated infectious bronchitis virus vaccine candidate derived from a Korean nephropathogenic strain

Vaccine ◽  
2010 ◽  
Vol 28 (16) ◽  
pp. 2887-2894 ◽  
Author(s):  
Hyun Jeong Lee ◽  
Ha Na Youn ◽  
Ji Sun Kwon ◽  
Youn Jeong Lee ◽  
Jae Hong Kim ◽  
...  
Keyword(s):  
Virus Vaccine ◽  
Infectious Bronchitis Virus ◽  
Vaccine Candidate ◽  
Infectious Bronchitis

scholarly journals A Real-Time PCR Method for the Authentication of Common Cuttlefish (Sepia officinalis) in Food Products

Foods ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 286 ◽  
Author(s):  
Amaya Velasco ◽  
Graciela Ramilo-Fernández ◽  
Carmen G. Sotelo
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Food Products ◽  
Polymerase Chain Reaction Method ◽  
Cross Reactivity ◽  
Sepia Officinalis ◽  
Base Pairs ◽  
Mitochondrial Coi ◽  
Fresh Frozen ◽  
Pcr Method

Cephalopods are very relevant food resources. The common cuttlefish (Sepia officinalis) is highly appreciated by consumers and there is a lack of rapid methods for its authentication in food products. We introduce a new minor groove binding (MGB) TaqMan real-time PCR (Polymerase Chain Reaction) method for the authentication of S. officinalis in food products to amplify a 122 base pairs (bp) fragment of the mitochondrial COI (Cytochrome Oxidase I) region. Reference and commercial samples of S. officinalis showed a threshold cycle (Ct) mean of 14.40, while the rest of the species examined did not amplify, or showed a significantly different Ct (p < 0.001). The calculated efficiency of the system was 101%, and the minimum DNA quantity detected was 10−4 ng. No cross-reactivity was detected with any other species, thus, the designed method differentiates S. officinalis from other species of the genus Sepia and other cephalopod species and works for fresh, frozen, grilled, cooked and canned samples of Sepia spp. The method has proved to be reliable and rapid, and it may prove to be a useful tool for the control of fraud in cuttlefish products.

scholarly journals Co-infection of bovine papillomavirus type-1 and -10 in teat warts of dairy cattle

2013 ◽  
Vol 58 (No. 12) ◽  
pp. 605-608
Author(s):  
P. Kumar ◽  
BL Jangir ◽  
G. Saikumar ◽  
R. Somvanshi
Keyword(s):  
Dairy Cattle ◽  
Real Time ◽  
Real Time Pcr ◽  
Rice Grain ◽  
Bovine Papillomavirus ◽  
Viral Dna ◽  
Specific Primers ◽  
Copy Numbers ◽  
Pcr Techniques

The present study was carried out to investigate the involvement of different bovine papillomaviruses in the teat warts of cattle. A total of 11 teat wart samples showing rice grain-like and small, sessile elevated greyish or flesh-like growths were collected from dairy cattle. DNA was extracted from these teat wart samples and PCR and real time PCR techniques were applied using specific primers for BPV-1 and -10 to detect the presence of viral nucleic acid. PCR revealed the presence of viral DNA of BPV-1 and -10 in three and seven samples, respectively. Quantification using real time PCR revealed that the copy numbers of the viral DNA of BPV-1 and -10 DNA varied from 1.12E + 04 to 2.99E + 04 and 3.56E + 02 to 5.23E + 06, respectively. From the present study it can be concluded that BPV-1 and -10 are involved in production of rice grain-like and sessile elevated growths on the teats of cattle.

Real-time PCR Assay with SNP-specific Primers for the Detection of a G143A Mutation Level in Venturia inaequalis Field Populations

2011 ◽  
Vol 159 (7-8) ◽  
pp. 569-578 ◽  
Author(s):  
Monika Michalecka ◽  
Tadeusz Malinowski ◽  
Agata Broniarek-Niemiec ◽  
Anna Bielenin
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Venturia Inaequalis ◽  
Pcr Assay ◽  
Specific Primers

scholarly journals Neutralization Assay for Zika and Dengue Viruses by Use of Real-Time-PCR-Based Endpoint Assessment

2017 ◽  
Vol 55 (10) ◽  
pp. 3104-3112 ◽  
Author(s):  
Heather L. Wilson ◽  
Thomas Tran ◽  
Julian Druce ◽  
Myrielle Dupont-Rouzeyrol ◽  
Michael Catton
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
High Sensitivity ◽  
Neutralization Assay ◽  
Cross Reactivity ◽  
Health Concern ◽  
Confirmatory Test ◽  
Virus Neutralization Assay ◽  
History Of ◽  
Infective Complications

ABSTRACTThe global spread and infective complications of Zika virus (ZKV) and dengue virus (DENV) have made them flaviviruses of public health concern. Serological diagnosis can be challenging due to antibody cross-reactivity, particularly in secondary flavivirus infections or when there is a history of flavivirus vaccination. The virus neutralization assay is considered to be the most specific assay for measurement of anti-flavivirus antibodies. This study describes an assay where the neutralization endpoint is measured by real-time PCR, providing results within 72 h. It demonstrated 100% sensitivity (24/24 ZKV and 15/15 DENV) and 100% specificity (11/11 specimens) when testing well-characterized sera. In addition, the assay was able to determine the correct DENV serotype in 91.7% of cases. The high sensitivity and specificity of the real-time PCR neutralization assay makes it suitable to use as a confirmatory test for sera that are reactive in commercial IgM/IgG enzyme immunoassays. Results are objective and the PCR-based measurement of the neutralization endpoint lends itself to automation so that throughput may be increased in times of high demand.

scholarly journals Rapid development and evaluation of a live-attenuated QX-like infectious bronchitis virus vaccine

Vaccine ◽  
2018 ◽  
Vol 36 (29) ◽  
pp. 4245-4254 ◽  
Author(s):  
Yun Zhang ◽  
Songjian Huang ◽  
Yuyao Zeng ◽  
Chunyi Xue ◽  
Yongchang Cao
Keyword(s):  
Virus Vaccine ◽  
Infectious Bronchitis Virus ◽  
Rapid Development ◽  
Infectious Bronchitis
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