scholarly journals Metapneumovirus infection of poultry: development of sovereign method of diagnosis and epizootological monitoring in poultry farms of Ukraine

2021 ◽  
pp. 9-14
Author(s):  
O.V. Tsinoviy ◽  
◽  
L.I. Nalyvayko ◽  
Keyword(s):  
Diagnostic System ◽  
Biological Properties ◽  
Test System ◽  
Chain Reaction ◽  
Subtype B ◽  
Epidemiological Monitoring ◽  
Poultry Farms ◽  
Indirect Hemagglutination ◽  
Polymerase Chain ◽  
Production Inspection

The main biological (cultural) properties of field isolates of metapneumovirus (MPV) were studied, normal and hyperimmune sera to metapneumovirus for IHR-diagnosticum (erythrocyte diagnosticum based on indirect hemagglutination reaction) were obtained, and a set of M-component components was developed. A diagnostic system of IHR (indirect hemagglutination reaction) was developed, with the help of which epizootological monitoring was carried out in poultry farms and the spread of a new infectious disease of poultry (metapneumovirus infection) among turkeys and chickens in poultry farms of Ukraine was studied. The purpose of the research is to develop a domestic diagnostic test system (erythrocyte diagnosticum for IHR) for metapneumovirus infection (MPVI) or infectious avian rhinotracheitis (IRT). As a result of the conducted researches metapneumovirus infection or infectious rhinotracheitis was established in poultry farms of 4 regions of Ukraine. The pathogen was isolated, its molecular-biological properties were studied by PCR (polymerase chain reaction), and it was established that it belongs to the genus Metapneumovirus (MPV), subtype B. The results of the epidemiological monitoring indicate that the developed erythrocyte MPV (IRT) antigen based on IHR is sensitive and specific and can be used to control the spread of metapneumovirus infection and the intensity of immunity in birds vaccinated against this disease. The production inspection in poultry farms in the western regions of Ukraine established the possibility of using IHR-diagnosticum for control of MPVI. As a result of the performed work the new domestic method of diagnostics, forecasting and protection of poultry against a metapneumovirus infection is offered. Prospects for further research are to use this erythrocyte diagnosticum based on the indirect hemagglutination reaction to monitor metapneumovirus infection of birds in poultry farms in Ukraine and determine the epizootic situation for this disease.

scholarly journals Virulence-Associated Gene Profiles of Avian Pathogenic Escherichia coli Isolated From Broilers With Colibacillosis: A Pilot Study in Iran

2019 ◽  
Vol 7 (1) ◽  
pp. 4-8
Author(s):  
Payam Haghighi Khoshkhoo ◽  
Hadi Pourtaghi ◽  
Gita Akbariazad ◽  
Saeed Mokhayeri
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Economic Losses ◽  
Avian Pathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Biochemical Tests ◽  
Gene Profiles ◽  
Pathogenic Escherichia Coli ◽  
Poultry Farms ◽  
Polymerase Chain

Background: Avian pathogenic Escherichia coli (APEC) causes economic losses in the chicken industry worldwide. Objective: In this study, virulence-associated gene profiles of APEC isolates were investigated by polymerase chain reaction (PCR). Materials and Methods: A total of 60 Escherichia coli isolates were collected from 60 colibacillosis cases from 30 broiler poultry farms in Alborz, Tehran, and Golestan provinces, Iran. After identification by biochemical tests, DNA was extracted by boiling method and 5 virulence-associated genes including: iutA, hlyF, iroN, ompT, and iss were detected by 2 multiplex PCR protocols. Results: Of the 60 APEC isolates, 26 (43.3%) isolates had at least three virulence genes from which 12 (20%) isolates were positive for all 5 virulence genes, whereas 34 (56.6%) carried no investigated virulence genes. Presence of iutA, hlyF, iroN, ompT, and iss genes in the APEC isolates were 17 (28.3%), 17 (28.3%), 24 (40%), 26 (43.3%), and 23 (38.3%), respectively. Conclusion: According to the results, four different virulence-associated gene profiles were seen in isolates, from which profile 1 with 12 (20%) isolates was predominant. These findings were in agreement with the previous reports.

scholarly journals Cell reservoirs in lymph nodes infected with HIV-1 subtype E differ from subtype B: identification by combined in situ polymerase chain reaction and immunohistochemistry

2005 ◽  
Vol 19 (2) ◽  
pp. 255-263 ◽  
Author(s):  
Lertlakana Bhoopat ◽  
Tat S Rithaporn ◽  
Surapan Khunamornpong ◽  
Tanin Bhoopat ◽  
Clive R Taylor ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lymph Nodes ◽  
Chain Reaction ◽  
Subtype B ◽  
Polymerase Chain ◽  
Hiv 1

scholarly journals TP53 signature diagnostic system using multiplex reverse transcription–polymerase chain reaction system enables prediction of prognosis of breast cancer patients

Breast Cancer ◽  
2021 ◽  
Author(s):  
Shin Takahashi ◽  
Takafumi Fukui ◽  
Tadashi Nomizu ◽  
Yoichiro Kakugawa ◽  
Fumisyoshi Fujishima ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Prognostic Significance ◽  
Reaction System ◽  
Diagnostic System ◽  
Wild Type ◽  
Chain Reaction ◽  
Polymerase Chain Reaction System ◽  
Polymerase Chain

Abstract Background TP53 status based on TP53 signature, a gene expression profile to determine the presence or absence of TP53 mutation, is an independent prognostic factor of breast cancer. The purpose of this study was to develop a simple diagnostic system for TP53 signature status. Methods We developed a multiplex reverse transcription–polymerase chain reaction system to determine TP53 status. Based on this system, prospectively collected 189 patients with stage I and II breast cancer were determined to have TP53 mutant signature or TP53 wild-type signature. The prognostic significance of the TP53 signature by the diagnostic system was analyzed. Results The diagnostic accuracy of TP53 status and reproducibility of this diagnosis system was confirmed. Using the diagnostic system, 89 patients were classified as TP53 mutant signature and the remaining 100 cases were classified as TP53 wild-type signature. Recurrence-free survival (RFS) among patients with TP53 mutant signature was significantly shorter than that among those with TP53 wild-type signature. On univariate and multivariate analyses, the TP53 signature status was an independent predictor of RFS. RFS among patients with TP53 mutant signature was significantly shorter than that among those with TP53 wild-type signature in a cohort of estrogen receptor-positive breast cancer. Although a difference was not significant, no recurrent cases was observed in TP53 wild-type signature group in triple negative breast cancer. Conclusion This simple and precise diagnostic system to determine TP53 signature status may help in prognostic assessment, therapeutic decision-making, and treatment optimization in patients with breast cancer.

scholarly journals Determining the Most Accurate and Early Form for Detection of Lyme Disease

2020 ◽  
Vol 3 (1) ◽  
Author(s):  
Samantha Zaremba
Keyword(s):  
Polymerase Chain Reaction ◽  
Lyme Disease ◽  
Diagnostic System ◽  
Cost Effective ◽  
Serological Testing ◽  
Chain Reaction ◽  
Early Form ◽  
Chronic Lyme Disease ◽  
Diagnostic Standards ◽  
Polymerase Chain

Lyme disease is the most common vector-based disease with over 300,000 new cases each year. The current diagnostic system for those with Early Lyme disease is only about 40% accurate. Since the severity of symptoms and probability of recurrence significantly increases as time progresses, there is a need to find a new alternative diagnostic system for Early Lyme disease. Many solutions to this problem are considered within this document, most notably: polymerase chain reaction detection, metabolic biosignature testing, and OspC targeting serological testing. After further analysis, metabolic biosignature testing is considered the best alternative since it yields the highest diagnosis sensitivity with around 89% accuracy. Additionally, metabolic biosignature testing is both cost effective and widely applicable since it does not require the Erythema migran rash to be used. Once in place, Lyme disease will be able to be diagnosed quicker, thus reducing the number of cases of Chronic Lyme disease which significantly reduces personal quality of life and is often costly. Keywords: Early Lyme disease, Metabolic Biosignature, Serological testing, Borrelia burgdorferi, Diagnostic Standards, Polymerase Chain Reaction

scholarly journals Biological and Genetic Characterization of Pod Pepper Vein Yellows Virus-Associated RNA From Capsicum frutescens in Wenshan, China

2021 ◽  
Vol 12 ◽  
Author(s):  
Jiejun Peng ◽  
Shan Bu ◽  
Yueyan Yin ◽  
Mengying Hua ◽  
Kuangjie Zhao ◽  
...  
Keyword(s):  
Yunnan Province ◽  
Genetic Characterization ◽  
Biological Properties ◽  
Capsicum Frutescens ◽  
Chain Reaction ◽  
Positive Sense ◽  
Polymerase Chain ◽  
Pepper Vein Yellows Virus ◽  
Generation Sequencing

Tombusvirus-like associated RNAs (tlaRNAs) are positive-sense single-stranded RNAs found in plants co-infected with some viruses of the genus Polerovirus. Pod pepper vein yellows virus (PoPeVYV) was recently reported as a new recombinant polerovirus causing interveinal yellowing, stunting, and leaf rolling in Capsicum frutescens plants at Wenshan city, Yunnan province, China. The complete genome sequence of its associated RNA has now been determined by next-generation sequencing and reverse transcription (RT) polymerase chain reaction (PCR). PoPeVYV-associated RNA (PoPeVYVaRNA) (GenBank Accession No. MW323470) has 2970 nucleotides and is closely related to other group II tlaRNAs, particularly tobacco bushy top disease-associated RNA (TBTDaRNA, GenBank Accession No. EF529625). In infection experiments on Nicotiana benthamiana and C. frutescens plants, synergism between PoPeVYVaRNA and PoPeVYV was demonstrated, leading to severe interveinal yellowing of leaves and stunting of plants. The results provide further information on the genetic and biological properties of the various agents associated with pepper vein yellows disease (PeVYD).

MUTATION IN THE FMO3 LOCUS AND ITS DISTRIBUTION IN A GROUP OF AYRSHIRE BREEDING BULLS USED IN FARMS OF THE KRASNODAR TERRITORY

2021 ◽  
pp. 10-15
Author(s):  
Н.В. КОВАЛЮК ◽  
Е.В. ШИРЯЕВА ◽  
Л.И. ЯКУШЕВА ◽  
Ю.Ю. ШАХНАЗАРОВА
Keyword(s):  
Polymerase Chain Reaction ◽  
Mutant Allele ◽  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Test System ◽  
Frequency Of Occurrence ◽  
Wild Type ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Рыбный привкус в коровьем молоке вызван наличием нонсенс-мутации (g.39523051C>T) в гене бычьего FMO3. Нами разработана тест-система для выявления FMO3- полиморфизма, основанная на полимеразной цепной реакции с последующим анализом полиморфизма длин фрагментов рестрикции с использованием эндонуклеазы TaqI. Фрагменты, которые амплифицировались с участка гена FMO3 «дикого» типа, расщеплялись эндонуклеазой TaqI  на 2 фрагмента: 136 и 99 пн. Фрагменты, амплифицированные с мутантного аллеля, сайта рестрикции не имели (их размер составлял  235 пн). Определена частота встречаемости носителей мутации в отечественной субпопуляции айрширского скота. Установлено, что среди айрширских быков-производителей (n=45), принадлежащих различным отечественным и зарубежным племорганизациям, частота встречаемости носителей мутации в гене FMO составила 9%. Учитывая, что выявленные носители мутации интенсивно используются и могут передавать эту аномалию значительному числу дочерей, генотипирование по локусу FMO3 должно стать обязательным для быков-производителей и групп быкопроизводящих коров племенных айрширских хозяйств. The fishy taste in cow's milk is caused by the presence of a nonsense mutation (g.39523051C>T) in the bovine FMO3 gene. We have developed a test system for detecting FMO3 polymorphism based on a polymerase chain reaction followed by analysis of restriction fragment length polymorphism using TaqI endonuclease. Fragments that were amplified from the wild-type FMO3 gene site were cleaved by TaqI endonuclease into 2 fragments: 136 and 99 bp. The fragments amplified from the mutant allele did not have a restriction site (their size was — 235 bp). The frequency of occurrence of mutation carriers in the domestic subpopulation of Ayrshire cattle was determined. It was found that among Ayrshire bulls (n=45) belonging to various domestic and foreign breeding organizations, the frequency of occurrence of carriers of the mutation in the FMO gene was 9%. Given that the identified carriers of the mutation are intensively used and can transmit this anomaly to a significant number of daughters, genotyping by the FMO3 locus should become mandatory for breeding bulls and groups of bull-producing cows of breeding Ayrshire farms.

BIOCHEMICAL PROPERTIES OF ACTINOBACILLUS PLEUROPNEUMONIAE MUSEUM STRAINS

2021 ◽  
pp. 58-62
Author(s):  
A. S. Andrusevich ◽  
E. L. Krasnikova ◽  
M. M. Misteyko ◽  
I. I. Strelchenya ◽  
M. S. Struk ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Veterinary Medicine ◽  
Actinobacillus Pleuropneumoniae ◽  
Biochemical Properties ◽  
Test System ◽  
Chain Reaction ◽  
Polymerase Chain

The article provides data on the biochemical properties of museum strains of Actinobacillus pleuropneumoni-ae. The belonging of the strains of Actinobacillus pleuropneumoniae was confirmed in the polymerase chain reaction using the developed RUE «Institute of experimental veterinary medicine nam. of S.N. Wyshelessky» test system for detecting the genome of Actinobacillus pleuropneumoniae.

Detection of Enteroviral RNA Sequences in Different Water Samples

1993 ◽  
Vol 27 (3-4) ◽  
pp. 219-222 ◽  
Author(s):  
D. Tougianidou ◽  
K. Botzenhart
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Water Samples ◽  
Test System ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rna Sequences ◽  
Pcr Product ◽  
Filter Unit ◽  
Polymerase Chain

Viruses were isolated from different water samples by Sterivex-Filtration. The nucleic acids were isolated in the filter unit and purified by phenol-chloroform extraction and ethanol precipitation. Reverse transcription and polymerase chain reaction (RT/PCR) were performed with primer pairs complementary to sequences of the enteroviral 5’ noncoding region. Amplified sequences were detected by hybridisation with an oligonucleotide complementary to a part of the PCR product. The test system seems to be sensitive and specific In the detection of enteroviral RNA.

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