scholarly journals Comparison of Reverse Transcriptase–Polymerase Chain Reaction, Virus Isolation, and Immunoperoxidase Assays for Detecting Pigs Infected with Low, Moderate, and High Virulent Strains of Classical Swine Fever Virus

2004 ◽  
Vol 16 (2) ◽  
pp. 132-138 ◽  
Author(s):  
Kathy Handel ◽  
Helen Kehler ◽  
Kevin Hills ◽  
John Pasick
1997 ◽  
Vol 13 (3) ◽  
pp. 239-244
Author(s):  
S. D. Kirilenko ◽  
O. M. Deriabin ◽  
O. G. Deriabina ◽  
O. L. Kirilenko ◽  
Ju. O. Cherednik ◽  
...  

1997 ◽  
Vol 9 (2) ◽  
pp. 118-124 ◽  
Author(s):  
G. Shad ◽  
W. C. Wilson ◽  
J. O. Mecham ◽  
J. F. Evermann

A reversible target capture viral RNA extraction procedure was combined with a reverse-transcriptase nested polymerase chain reaction (PCR) to develop a capture PCR assay providing a rapid and safe prediction method for circulating bluetongue virus in infected ruminants. This new assay was compared with virus isolation and a recently developed antigen-capture enzyme-linked immunosorbent assay (ELISA) for the detection of bluetongue virus. Eight Warhill crossbred sheep were inoculated subcutaneously with bluetongue virus serotype 10, and blood samples were taken sequentially over a period of 28 days. The capture PCR detected the peak of viremia, as determined by virus isolation and antigen-capture ELISA, from day 5 to day 14 after challenge. The results indicate that the rapid-capture bluetongue virus PCR provides a rapid indicator of samples in which virus can be isolated. In addition, this capture bluetongue virus PCR procedure does not require a lengthy phenol extraction or the use of the highly toxic methyl mercury hydroxide denaturant.


Author(s):  
Emmanuel G. Kifaro ◽  
Japhet Nkangaga ◽  
Gradson Joshua ◽  
Raphael Sallu ◽  
Mmeta Yongolo ◽  
...  

Rift Valley fever virus (RVFV) is an acute, zoonotic viral disease caused by a  Phlebovirus, which belongs to the Bunyaviridae family. Among livestock, outbreaks of the disease are economically devastating. They are often characterised by large, sweeping abortion storms and have significant mortality in adult livestock. The aim of the current study was to investigate RVFV infection in the Kigoma region, which is nestled under the hills of the western arm of the Great Rift Valley on the edge of Lake Tanganyika, Tanzania. A region-wide serosurvey was conducted on non-vaccinated small ruminants (sheep and goats, n = 411). Sera samples were tested for the presence of anti-RVFV antibodies and viral antigen, using commercial enzyme-linked immunosorbent assay and reverse transcriptase polymerase chain reaction, respectively. The overall past infections were detected in 22 of the 411 animals, 5.4% (Confidence Interval (CI) 95% = 3.5% – 8.1%). The Kigoma rural area recorded the higher seroprevalence of 12.0% (CI 95% = 7.3% – 18.3%; p < 0.0001), followed by Kibondo at 2.3% (CI 95% = 0.5% – 6.5%; p > 0.05) and the Kasulu district at 0.8% (CI 95% = 0.0% – 4.2%; p > 0.05). The prevalence was 12.5% and 4.7% for sheep and goats, respectively. Reverse transcriptase polymerase chain reaction results indicated that only eight samples were found to be positive (n = 63). This study has confirmed, for the first time, the presence of the RVFV in the Kigoma region four years after the 2007 epizootic in Tanzania. The study further suggests that the virus activity exists during the inter-epizootic period, even in regions with no history of RVFV.


2003 ◽  
Vol 15 (1) ◽  
pp. 35-41 ◽  
Author(s):  
Changsun Choi ◽  
Chanhee Chae

A seminested reverse transcription–polymerase chain reaction (RT-PCR) was developed for the detection of classical swine fever virus (CSFV) in semen. Five boars were inoculated intranasally with CSFV isolate propagated in PK15 cells. Two boars inoculated with the supernatant of noninfected PK15 cells were kept as controls. Semen and serum samples were collected twice weekly for 63 days postinoculation (dpi). Samples were tested for the presence of antibodies to CSFV by an enzyme-linked immunosorbent assay and for the presence of CSFV nucleic acid by seminested RT-PCR. Antibodies to CSFV could be detected as early as 7 dpi in 1 boar, and all 5 infected boars were found positive by 14 dpi. CSFV from boar semen was infrequently identified by virus isolation compared with seminested RT-PCR. CSFV nucleic acid was detected in semen by seminested RT-PCR as early as 7 dpi in 3 infected boars and persistently thereafter in all 5 infected boars until 63 dpi. When separated fractions of CSFV-contaminated semen were analyzed by the seminested RT-PCR, the CSFV nucleic acid was detected mainly in seminal fluid and occasionally in nonsperm cells. CSFV antigen was also detected in nonsperm cells from semen smear by immunohistochemistry. Thus, infection via semen, specially through CSFV-infected seminal fluid, seems to be a major route of transmission of CSFV.


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