[Establishment of in situ hybridization technique for detection of parvovirus B19 and its clinical application].
OBJECTIVE
To establish in situ hybridization (ISH) technique to detect parvovirus B19 and to know the viral location in cardiac tissue of congenital heart diseases (CHD).


METHODS
ISH method was established with randomly primed, degoxigenin labeled B19 DNA probe of 1112 bp long corresponding to parvovirus B19 capsid protein VP1 genome.


RESULTS
Dot blot hybridization showed that B19 DNA was positive with the sensitivity of 0.1 pg/microl, while CMV, HSV, TOX DNA were all negative. 7 out of 66 CHD cardiac tissues were B19 DNA positive by using ISH, while in 38 cases of control group were all negative. B19 DNA positive signal was found in the nucleus of cardiac cell.


CONCLUSIONS
We have established ISH to detect B19 DNA that has high sensitivity (0. 1 pg/microl) and specificity and have found that B19 DNA is located the nucleus of cardiac cell.
Wang
X
X
Department of Pediatrics, Xijing Hospital of the Fourth Military Medical University, Xian 710032, China.
Zhang
G
G
Li
Y
Y
D000293:Adolescent; D000328:Adult; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D006330:Heart Defects, Congenital / Q000821:virology*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D016732:Parvovirus B19, Human / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D004279:DNA, Viral