Development and evaluation of a sensitive PCR-ELISA for detection of adenoviruses in feces.
OBJECTIVE
Most of the PCR-based assays for the detection of adenoviruses require gel electrophoresis for resolution, which is time- and work-consuming and interpretation difficulties often occur. Therefore our objective was to develop and evaluate a PCR-ELISA for broad detection of adenoviruses, that is easy to perform and leads to an increase of specificity in virus detection.


METHODS
A total of 64 fecal specimens from patients with acute gastroenteritis, previously tested negative for other viruses than adenovirus and for bacterial pathogens, were tested for adenovirus in parallel with an Antigen-ELISA, virus isolation in cell culture, with a previously published hybridization-based PCR and our newly developed semi-nested PCR-ELISA.


RESULTS
The new PCR-ELISA turned out to be the best method with 26 samples positive for adenovirus. Of these, 22 became positive by Ag-ELISA, 19 by virus isolation in cell culture and only 14 by the previously published hybridization-based PCR.


CONCLUSIONS
The semi-nested PCR-ELISA is a sensitive, reliable and rapid method for the detection of enteric adenoviruses from fecal samples.
Metzger-Boddien
Christoph
C
AnDiaTec GmbH & Co. KG and MB-Diagnostik GmbH, Kornwestheim, Germany.
Kehle
Johannes
J
D000258:Adenovirus Infections, Human / Q000175:diagnosis*; D000260:Adenoviruses, Human / Q000302:isolation & purification*; D000956:Antigens, Viral / Q000032:analysis; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000032:analysis / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005145:Face / Q000821:virology*; D006801:Humans; D007223:Infant; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014776:Virus Cultivation
D000956:Antigens, Viral; D004279:DNA, Viral