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					                                                                                   Bug                          BITES
               ISSUE No. 35  June 2007                                                                                     LAB NEWS AND VIEWS
                                                                                                 emerging PRRSV variants. In July of 2007, we will
                                                                                                 launch a second generation PRRSV test that utilizes
                                                                                                 new NA PRRSV primers and probe designed by Steven
                                                                                                 Kleiboeker DVM, PhD, University of Missouri, based on
                                                                                                 current sequencing information. These new primers and
                                                                                                 probe target highly conserved regions in the 3’-
                                                                                                 untranslated region (UTR) of the NA PRRS virus.
                                                                                                 Before initiating the process to replace the current test,
                                                                                                 research efforts confirmed that the new NA primers and
                                                                                                 probe could be successfully multiplexed with the current
                                                                                                 EU primers and probe without sacrificing sensitivity and
                                                                                                 specificity. So, the simultaneous detection and
                                                                                                 discrimination of the NA and EU strains and their
                                                                                                 detection limits remain unchanged.
                                                                                                 Validation of the new real-time 3’-UTR NA/EU multiplex
                                                                                                 PRRSV PCR test was performed in parallel with our
BI PRRSV Testing Proficiency Panels                                                              current PRRSV PCR test on 116 field samples. The
The PRRSV proficiency panel was established by                                                   data was compared to assure that the performance of
Boehringer Ingelheim (BI) and is provided free of charge                                         the new test was equivalent or superior to the current
to interested laboratories. It is an unbiased way to                                             test offered by Gallant. The validation data
assess the testing abilities of laboratories conducting                                          demonstrated that the new 3’-UTR PRRSV real-time
PCR tests to detect PRRS virus. BI submits blinded                                               PCR test had excellent correlation (97.4%) with
samples of virus (diluted to produce both positives and                                          Gallant’s original PRRSV real-time PCR test.
negatives) to participating laboratories. The results are                                        In addition, analysis of PRRSV data collected over a
reported to BI who analyze the data and provide an                                               one year period identified 5% of the farms/systems
anonymous comparative data table to participants.                                                tested at Gallant had PRRSV variants that were
Gallant Labs participated in the September 2006 and                                              detected more efficiently with the new 3’-UTR NA/EU
March 2007 BI proficiency panels with excellent results.                                         multiplex PRRSV real-time PCR test. This suggests
Comparison of the PRRSV proficiency data positions                                               that the new 3’-UTR directed primers and probe would
Gallant among the labs exhibiting high sensitivity without                                       offer increased sensitivity, detection and quantification
producing false positives. In addition, Gallant’s virus                                          of emerging NA PRRSV variants in the clinical samples
quantitation results are in agreement with other real-time                                       analyzed.
PCR testing labs that reported virus titers.
                                                                                                 Benefits of the new PRRSV multiplex assay to
Next Generation Real Time RT-PCR Test for                                                        Clients:
PRRSV Launch By Gallant Custom Laboratories
                                                                                                      Improved detection of all NA PRRSV strains
One of the challenges for both control and testing of
                                                                                                       including emerging NA PRRSV variants.
PRRS is the continual genetic mutation of the PRRS
virus. To address this concern, Gallant continues to                                                  More accurate estimation of NA PRRS virus titer.
make molecular improvements to our North American                                                     Detection limit is unchanged and remains at 5 virus
(NA) and European (EU) multiplex PRRSV real-time                                                       copies or better.
PCR test to ensure that the test we provide is sensitive                                              Both NA and EU PRRS viruses can be simultaneous
and minimizes the possibility of false negatives due to                                                detected, identified and quantified in one test.

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