scholarly journals Prevalence of canine coronavirus and parvovirus infections in dogs with gastroenteritis in Thailand

2012 ◽  
Vol 48 (No. 6) ◽  
pp. 163-168 ◽  
Author(s):  
K. Sakulwira ◽  
P. Vanapongtipagorn ◽  
A. Theamboonlers ◽  
K. Oraveerakul ◽  
Y. Poovorawan
Keyword(s):  
Polymerase Chain Reaction ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Gene Encoding ◽  
Nested Polymerase Chain Reaction ◽  
Fecal Samples ◽  
Canine Coronavirus ◽  
Pcr Product ◽  
Causative Agents ◽  
Polymerase Chain

Canine coronavirus (CCV) and canine parvovirus type 2 (CPV-2) are the causative agents of gastroenteritis in dogs. Seventy fecal samples from dogs with signs of gastroenteritis (vomiting and diarrhea), twenty-five fecal samples from healthy dogs and one CPV-2 vaccine strain were amplified by semi-nested polymerase chain reaction (PCR) and semi-nested reverse transcriptase polymerase chain reaction (RT-PCR), aimed at specifically studying the gene encoding the most abundant capsid protein VP2 of CPV-2 and spike protein of CCV. The specificity of the CCV RT-PCR product was evaluated by sequencing. Positive specimens comprised 44 samples (62.8%) and 9 samples (12.8%) for CPV-2 and CCV, respectively. In nine CCV positive samples, seven displayed co-infection between CCV and CPV-2. Our CCV sequence (AF482001) showed a 94.9% nucleotide identity to CCV reported in GenBank accession number D13096. High prevalence of CCV and CPV-2 infections was found in 1–2 month- and 3–6 month-old dogs, respectively. Molecular biology of these viruses is important primarily for epidemic control and preventive measures.

scholarly journals Pet rat harbouring Seoul hantavirus in Sweden, June 2013

2013 ◽  
Vol 18 (27) ◽  
Author(s):  
Å Lundkvist ◽  
J Verner-Carlsson ◽  
A Plyusnina ◽  
L Forslund ◽  
R Feinstein ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lung Tissue ◽  
Reverse Transcription ◽  
Haemorrhagic Fever ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Specific Antibodies ◽  
Polymerase Chain ◽  
Focus Reduction

We report the first detection of Seoul hantavirus (SEOV) in a pet rat in Sweden. SEOV-specific antibodies were detected in the pet rat blood by focus reduction neutralising test (FRNT), and SEOV RNA in lung tissue was confirmed by reverse transcription-nested polymerase chain reaction (RT-PCR) followed by sequencing. The discovery follows the recent reports of SEOV infected pet rats, as well as associated human cases of severe haemorrhagic fever with renal syndrome (HFRS), in England and Wales.

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.

scholarly journals Detection of Porcine Circovirus Type 2 in Feces of Pigs with or without Enteric Disease by Polymerase Chain Reaction

2003 ◽  
Vol 15 (4) ◽  
pp. 369-373 ◽  
Author(s):  
Jeong S. Yang ◽  
Dae S. Song ◽  
So Y. Kim ◽  
Kwang S. Lyoo ◽  
Bong K. Park
Keyword(s):  
Polymerase Chain Reaction ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Porcine Circovirus ◽  
Enteric Disease ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Fecal Samples ◽  
Polymerase Chain

To establish the sensitive polymerase chain reaction(PCR) method and detect porcine circovirus type 2 (PCV2) from intestines and feces of commercial swine herds with or without enteric disease, intestinal samples from 68 pigs and 29 fecal samples from commercial swine farms were collected. A primer set, forward primer 5′-GAAGAATGGAAGAAGCGG-3′ and reverse primer 5′-CTCACAGCAGTAGACAGGT-3′, could detect the virus at a concentration as low as 2 infectious virions per milliliter under controlled conditions using PK-15 cell-adapted PCV2. The genomic nucleotide sequences of open reading frame 1 (ORF1) PCR products from fecal samples were found to have complete homology with other PCV2s deposited in the GenBank database. Transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) as the other enteric pathogens were also investigated by performing duplex reverse transcription-PCR (RT-PCR). Among 63 pigs with clinical enteric disease, 18 PCV2s (14 from intestines and 4 from feces), 7 TGEVs from intestines, and 18 PEDVs (14 from intestines and 4 from feces) were detected by PCR and the duplex RT-PCR. In 34 pigs (14 from intestines and 20 from feces) without clinical enteric disease, only PCV2 was detected in 19 pigs (3 from intestines and 16 from feces). Both PEDV and PCV2 were found in 6 pigs with clinical enteric disease. Among 15 PCV2 samples that were PCR-positive, 4 were culture-positive at passage level 3 in PK-15 cells. These results reveal that PCV2 is shed through the feces of pigs without clinical enteric disease, which suggests the potentiality of the fecal–oral transmission of PCV2 in feces.

scholarly journals The Epidemiological Characteristics of the Korean Bat Paramyxovirus between 2016 and 2019

2020 ◽  
Vol 8 (6) ◽  
pp. 844
Author(s):  
Seong Sik Jang ◽  
Ji Yeong Noh ◽  
Van Thi Lo ◽  
Yong Gun Choi ◽  
Sun-Woo Yoon ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Nested Pcr ◽  
Human Pathogens ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Rdrp Region ◽  
Polymerase Chain ◽  
Epidemiological Characteristics

Bats are considered reservoirs of severe emerging human pathogens. Notably, bats host major mammalian paramyxoviruses from the family Paramyxoviridae, order Mononegavirales. In this study, paramyxoviruses were investigated by reverse transcription semi-nested polymerase chain reaction (RT-semi-nested PCR) and reverse transcription polymerase chain reaction (RT-PCR), based on the RT-semi-nested PCR using the consensus paramyxovirus primers targeting the RNA dependent-RNA-polymerase (RdRp) region. In addition, RT-PCR was performed using newly designed primers targeting regions of the fusion protein (F) and hemagglutinin-neuraminidase (HN). The dominant bat species in the collection site of paramyxoviruses were Miniopterus schreibersii, Myotis macrodactylus, Myotis petax, and Rhinolophus ferrumequinum. Paramyxoviruses were detected in four samples in 2016 and six in 2019. Meanwhile, in samples collected in 2017 and 2018, no paramyxoviruses were detected. Phylogenetic analysis based on the partial nucleotide sequences of RdRp, F, and HN proteins suggested that the viruses belonged to the proposed genus Shaanvirus. In conclusion, this study revealed that bat paramyxoviruses in Korea belonged to a single genus and circulated sporadically in several provinces, including Chungbuk, Gangwon, Jeju, and Jeonnam.

scholarly journals OCCURRENCE OF CRYPTOSPORIDIUM SP. IN DOGS AND CATS FROM CURITIBA AND ITS METROPOLITAN AREA

2013 ◽  
Vol 18 (3) ◽  
Author(s):  
Martha Grecca ◽  
Vanete Thomaz-Soccol ◽  
Magda C Costa Ribeiro ◽  
Jessé Henrique Truppel ◽  
Juliana Tracz Pereira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Infection Rate ◽  
Metropolitan Area ◽  
Epidemiological Data ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Fecal Samples ◽  
Rdna Gene ◽  
Polymerase Chain ◽  
Low Sensitivity

The present study was carried out with the aim of assess the occurrence of Cryptosporidium sp. infection in dogs and cats in Curitiba and its metropolitan area, state of Paraná, Brazil. Techniques employed to detect the protozoan in fecal samples were: staining by Ziehl-Neelsen for oocysts search and nested polymerase chain reaction (nPCR) targeting the 18SSU rDNA gene. To attempt the proposed aim, 91 feces samples of dogs and 25 of cats were collected and analyzed. Ziehl-Neelsen technique was unable to detect any oocyst in both groups analyzed, showing a very low sensitivity. Results of nPCR showed an infection rate of 13.2% (12/91) and 4% (1/25) in dogs and cats respectively.  The implications of these epidemiological data are discussed in this work.

scholarly journals Detection of mouse hepatitis virus in mouse colonies using the nested polymerase chain reaction

2000 ◽  
Vol 52 (4) ◽  
pp. 307-312
Author(s):  
A.B. Cecílio ◽  
A.L. Cândido ◽  
M. Resende ◽  
E.D. Bontempo ◽  
A.S. Martins
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Hepatitis Virus ◽  
Mouse Hepatitis Virus ◽  
Hepatic Tissue ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Specific Amplification ◽  
Polymerase Chain

A reverse transcriptase polymerase chain reaction (RT-PCR) to detect mouse hepatitis virus (MHV) in hepatic tissue was developed. To circumvent possible failures in RT-PCR amplifications, a second round of PCR with internal primers was used to confirm the specificity and increase the sensitivity of the test. Using this method specific amplification of MHV sequences was observed in 18 out of 20 mouse colonies examined.

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