Abstract
An improved multiplex PCR, using redesigned primers targeting the serovar 3 capsule locus, which differentiates serovars 3, 6, and 8 Actinobacillus pleuropneumoniae isolates, is described. The new primers eliminate an aberrant serovar 3-indicative amplicon found in some serovar 6 clinical isolates. Furthermore, we have developed a new multiplex PCR for the detection of serovars 1 to 3, 5 to 8, 10, and 12 along with apxIV, thus extending the utility of this diagnostic PCR to cover a broader range of isolates.
Highlights
An improved multiplex PCR, using redesigned primers targeting the serovar 3 capsule locus, which differentiates serovars 3, 6, and 8 Actinobacillus pleuropneumoniae isolates, is described
We have devised a multiplex PCR that determines whether an isolate is serovar 3, 6, or 8 [10]. The latter multiplex PCR amplifies a fragment of the A. pleuropneumoniae-specific apxIV gene and serovar 3, 6, and 8-specific sequences derived from the capsule loci
We have observed in our collection a double- banding pattern in some isolates, so that it is not possible to distinguish between serovars 3 and 6
Summary
An improved multiplex PCR, using redesigned primers targeting the serovar 3 capsule locus, which differentiates serovars 3, 6, and 8 Actinobacillus pleuropneumoniae isolates, is described. We (i) discerned the genetic reason for the aberrant serovar 3/6 double-banding pattern, (ii) developed a new serovar 3-6-8 PCR with modified serovar 3 primers that eliminate the aberrant pattern, and (iii) extended that PCR to include a further six serovars commonly found throughout the world. This includes the detection of serovar 10, for which a capsule locus-based PCR has not been described previously
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