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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #84977
Title: RT-PCR SENSITIVITY COMPARISON OF ORF 3-7 OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS

Author
item Umthun, Angela
item Mengeling, William

Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 11/10/1997
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped, positive sense RNA virus of the arteriviridae family. PRRSV infections are characterized by reproductive failure in gilts and sows and respiratory distress in pigs of all ages. A fast and efficient method for detecting PRRSV is by RT-PCR. We report here a RT-PCR sensitivity comparison for open reading frames (ORFs) 3-7. Primers were designed on either side of each ORF. Identical RT-PCR conditions were used for a single round of amplification of each ORF. The RT-PCR sensitivity for each ORF was determined using the same serial dilutions of total RNA purified from MARC-145 cells infected with either a wild-type or tissue culture adapted version of PRRSV isolate NADC 9 (passage 2 or 252 respectively). When wild-type virus was used the minimum detectable dilution was 10**-3 for each ORF, which is equivalent to 1.4 x 10**3 infectious units/ml. When NADC 9 (252) was used the minimum detectable dilution was 10**-4 to 10**-5 for each ORF, equivalent to approximately 1 x 10**3 infectious units/ml. Although it might be expected that ORF 7 would be the most sensitive template , our results suggest that ORFs 3-7 are equally sensitive templates for the detection of PRRSV by RT-PCR.