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PCR detection of Actinobacillus pleuropneumoniae apx IV gene in formalin‐fixed, paraffin‐embedded lung tissues and comparison with in situ hybridization
Author(s) -
Cho W.S.,
Chae C.
Publication year - 2003
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2003.01347.x
Subject(s) - actinobacillus pleuropneumoniae , biology , serotype , in situ hybridization , nested polymerase chain reaction , lung , polymerase chain reaction , microbiology and biotechnology , gene , pathology , gene expression , medicine , genetics
Aims: 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 apx IV gene. Methods and Results: The PCR results from formalin‐fixed, paraffin‐embedded tissues were compared with in situ hybridization. The apx IV 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 apx IV 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 apx IV 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 apx IV gene to diagnose A. pleuropneumoniae infection when formalin‐fixed tissues are submitted. Significance and Impact of the Study: This study confirmed the possibility of using formalin‐fixed, paraffin‐embedded tissues for the diagnosis of A. pleuropneumoniae infection in pigs.