Analysing myocardial tissue from explanted hearts of heart transplant recipients and multi-organ donors for the presence of parvovirus B19 DNA.
BACKGROUND
Parvovirus B19 (PVB19) is an erythrovirus causing diverse clinical manifestations ranging from asymptomatic or mild to more severe outcomes, dependent on the haematological and/or immunological status of the host. Reports of PVB19 infection as a causative agent of paediatric or adult inflammatory cardiac diseases, or of cardiac transplant rejection are rare.


OBJECTIVE
To identify PVB19 and other cardiotropic viruses in the myocardium of heart transplant (HTx) recipients and multi-organ donors (MOD). Furthermore, to assess the prevalence of cardiotropic viral infection in inflammatory heart disease.


METHODS
Heart tissue samples from 110 explants were analysed for PVB19 using primers and a 5'-nuclease probe designed to amplify a 160-basepair PCR product from the VP1/NS1 gene region. Samples tested included those obtained from patients undergoing HTx or from MODs. The findings were correlated with clinical course, histologic analysis and serologic testing. Confirmation of the positive PCR-results was done by sequencing and in situ hybridisation.


RESULTS
The new assay described here allows precise quantitation of viral load over 7 orders of magnitude (10(6) to 10(0)IU/assay). Measurable amounts of parvoviral genomes were detected in 4/56 (7%) explanted HTx-hearts and in 5/54 (9%) explanted MOD-hearts.


CONCLUSIONS
The newly developed real-time PCR is a rapid, sensitive and specific method to detect PVB19 infection in heart tissue. It will be a useful tool to address important questions regarding viral infections transmitted by transplantation, acute infections, relapses and complications involving late or chronic rejection.
Robert Koch-Institut, Nordufer 20, D-13353 Berlin, Germany. donosoo@rki.de
Oliver
O
Donoso Mantke
Andreas
A
Nitsche
Rudolf
R
Meyer
Karin
K
Klingel
Matthias
M
Niedrig
D000293:Adolescent; D000328:Adult; D000368:Aged; D000914:Antibodies, Viral / Q000097:blood; D036022:Capsid Proteins / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D006321:Heart / Q000821:virology*; D016027:Heart Transplantation; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D008875:Middle Aged; D010322:Parvoviridae Infections / Q000821:virology*; D016732:Parvovirus B19, Human / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014019:Tissue Donors; D017361:Viral Nonstructural Proteins / Q000235:genetics
D000914:Antibodies, Viral; D036022:Capsid Proteins; D004279:DNA, Viral; C106202:NS1 protein, parvovirus; D017361:Viral Nonstructural Proteins; C095547:capsid protein VP1, parvovirus B19