Development of polymerase chain reaction and comparison with in situ hybridization for the detection of Haemophilus parasuis in formalin-fixed, paraffin-embedded tissues.
DNA extraction and nested polymerase chain reaction (PCR) were developed for the detection of Haemophilus parasuis from formalin-fixed, paraffin-embedded tissues. The results for nested PCR were compared with those determined by in situ hybridization. The optimal results obtained show that use of xylene deparaffinization, digestion with proteinase K followed by nested PCR is a reliable detection method. A distinct positive signal was detected in 20 pigs naturally infected with H. parasuis by in situ hybridization. The rate of agreement between nested PCR and in situ hybridization for the detection of H. parasuis in formalin-fixed, paraffin-embedded tissues was 100%. The nested PCR could be applied successfully to formalin-fixed, paraffin-embedded tissues for the detection of H. parasuis with bacterial isolation.
Department of Veterinary Pathology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Republic of Korea.
Kwonil
K
Jung
Yoonchul
Y
Ha
Sung-Hoon
SH
Kim
Chanhee
C
Chae
D016977:Actinobacillus pleuropneumoniae / Q000235:genetics; D000818:Animals; D003126:Colostrum; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis; D005557:Formaldehyde; D006192:Haemophilus Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D044137:Haemophilus parasuis / Q000235:genetics / Q000302:isolation & purification*; D017403:In Situ Hybridization / Q000379:methods / Q000662:veterinary*; D008099:Liver / Q000382:microbiology; D016612:Paraffin Embedding / Q000662:veterinary; D016979:Pasteurella multocida / Q000235:genetics; D010496:Pericardium / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013154:Spleen / Q000382:microbiology; D013552:Swine / Q000382:microbiology; D013553:Swine Diseases / Q000175:diagnosis / Q000382:microbiology*; D016707:Tissue Fixation / Q000662:veterinary
D017931:DNA Primers; D004269:DNA, Bacterial; D005557:Formaldehyde