Detection and distribution of DNA of Actinobacillus pleuropneumoniae in the lungs of naturally infected pigs by in-situ hybridization.
Detection and distribution of Actinobacillus pleuropneumoniae was studied in formalin-fixed paraffin wax-embedded lung tissues from 10 naturally infected pigs by in-situ hybridization with a non-radioactive digoxigenin-labelled probe. A 610 base pair cDNA probe from a genomic library of A. pleuropneumoniae was generated by the polymerase chain reaction. All 10 pigs infected with A. pleuropneumoniae serotypes 2, 5, 6, or an untypable strain showed a distinct, positive signal in the degenerate alveolar leucocytes bordering a zone of coagulative necrosis and in the dense zone of degenerated cells in granulation tissue surrounding the necrotic areas. Positive cells typically exhibited dark-brown to black labelled deposits without background staining. A. pleuropneumoniae nucleic acids were more readily detected in areas of coagulative necrosis than in areas of granulation tissue. In-situ hybridization demonstrated that A. pleuropneumoniae primarily infected neutrophils and alveolar macrophages. The technique used was rapid, specific and sensitive, and may prove useful for the diagnosis of A. pleuropneumoniae infection in routinely fixed and processed tissues, obviating the need for bacterial isolation.
Min
K
K
Department of Veterinary Pathology, College of Veterinary Medicine, Seoul National University, Kyounggi-Do, Republic of Korea.
Chae
C
C
D000189:Actinobacillus Infections / Q000382:microbiology / Q000473:pathology / Q000662:veterinary*; D016977:Actinobacillus pleuropneumoniae / Q000235:genetics*; D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis*; D017403:In Situ Hybridization / Q000662:veterinary*; D008168:Lung / Q000737:chemistry / Q000382:microbiology / Q000473:pathology; D016612:Paraffin Embedding; D018410:Pneumonia, Bacterial / Q000382:microbiology / Q000473:pathology / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases / Q000382:microbiology* / Q000473:pathology
D017931:DNA Primers; D004269:DNA, Bacterial