Development of in-situ hybridization for the detection of Mycoplasma haemosuis (Eperythrozoon suis) in formalin-fixed, paraffin wax-embedded tissues from experimentally infected splenectomized pigs.
Mycoplasma haemosuis DNA was detected in experimentally infected splenectomized pigs by in-situ hybridization (ISH) with a nonradioactive digoxigenin-labelled DNA probe. An 839 base pair DNA probe targeting a 16S rRNA gene was generated by the polymerase chain reaction. Eight 6-week-old pigs were inoculated intraperitoneally with 6 ml of M. haemosuis-infected pig blood and eight negative control pigs were inoculated intraperitoneally with 6 ml of M. haemosuis-free blood. Two pigs from each group were killed for examination at 3, 7, 15 and 30 days post-inoculation (dpi). Red blood cells infected with M. haemosuis were first detected by light microscopy at 3 to 7 dpi. No M. haemosuis was observed in negative control pigs. Hybridization signals were evident in blood from the infected pigs at 3 dpi. The ISH method developed in this study was useful for the detection of M. haemosuis DNA in formalin-fixed, paraffin wax-embedded tissues and may be valuable for studying the pathogenesis of M. haemosuis infection.
Ha
S-K
SK
Departmentof Veterinary Parhology, College of Veterinary Medicine and School of Agricultural Biotechnology, Seoul National University, Seoul.
Jung
K
K
Choi
C
C
Ha
Y
Y
Song
H-C
HC
Lim
J-H
JH
Kim
S-H
SH
Chae
C
C
D000818:Animals; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis; D004912:Erythrocytes / Q000737:chemistry / Q000382:microbiology; D005557:Formaldehyde; D017403:In Situ Hybridization / Q000379:methods*; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D013156:Splenectomy; D013552:Swine; D013553:Swine Diseases / Q000235:genetics / Q000382:microbiology*
D015342:DNA Probes; D004269:DNA, Bacterial; D005557:Formaldehyde