PCR detection of Actinobacillus pleuropneumoniaeapxIV gene in formalin-fixed, paraffin-embedded lung tissues and comparison with in situ hybridization.
OBJECTIVE
Formalin-fixed, paraffin-embedded lung tissues from pigs experimentally infected with 12 Actinobacillus pleuropneumoniae serotypes were used to develop nested PCR for the detection of apxIV gene.


RESULTS
The PCR results from formalin-fixed, paraffin-embedded tissues were compared with in situ hybridization. The apxIV gene was detected in formalin-fixed, paraffin-embedded lung tissues from all 39 pigs experimentally infected with 12 A. pleuropneumoniae serotypes by nested PCR. In situ hybridization produced a distinct positive signal in all 39 pigs experimentally infected with 12 A. pleuropneumoniae serotypes. Agreement rates between nested PCR and in situ hybridization were 100% for the detection of apxIV gene in formalin-fixed paraffin-embedded lung tissues. Acceptable PCR signals were detected from lung tissues fixed for periods up to 180 days.


CONCLUSIONS
The apxIV gene is species-specific rather than serotype-specific and is therefore an important diagnostic marker. The nested PCR assay would be a useful method for the detection of apxIV gene to diagnose A. pleuropneumoniae infection when formalin-fixed tissues are submitted.


CONCLUSIONS
This study confirmed the possibility of using formalin-fixed, paraffin-embedded tissues for the diagnosis of A. pleuropneumoniae infection in pigs.
Cho
W-S
WS
Department of Veterinary Pathology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul, Republic of Korea.
Chae
C
C
D016977:Actinobacillus pleuropneumoniae / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis; D005404:Fixatives / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry*; D017403:In Situ Hybridization / Q000379:methods; D008168:Lung / Q000166:cytology / Q000382:microbiology*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D034421:Sus scrofa / Q000382:microbiology*; D013997:Time Factors
C122177:ApxIVA protein, Actinobacillus pleuropneumoniae; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005404:Fixatives; D005557:Formaldehyde