title: Development of duplex real-time PCR for detection of two DNA respiratory viruses.
abstract: A method was developed for the detection and quantitation of HAdV (human adenovirus) and HBoV (human bocavirus) based on a duplex real-time PCR, the AB PCR, using a Smartcycler instrument. A control real-time PCR was carried out on albumin DNA to standardise the non-homogenous respiratory samples. No cross-reactivity was observed with viruses or bacteria that could be found in the respiratory tract. The diagnosis rate using the AB PCR on clinical samples was 10.7%: 3.4% for HBoV detection, 6.9% for HAdV detection and 0.3% double detection HBoV-HAdV. The clinical and epidemiological characteristics of the HAdV- and HBoV-infected patients were evaluated. In the HAdV-positive group and the HBoV-positive group the samples were classified according to the severity of the disease. The HAdV viral load did not appear to be linked to the severity of the disease. Conversely, the difference between the two HBoV groups, severe and non-severe, was significant statistically when the comparison was based on the viral load (P=0.006) or after adjustment of the viral load to the number of cells in the samples (P=0.02).
authors: Department of Virology, University Hospital of Caen, Avenue Georges Clemenceau, 14033 Caen Cedex, France. dina-j@chu-caen.fr
authors: Julia
authors: J
authors: Dina
authors: Emilie
authors: E
authors: Nguyen
authors: Stephanie
authors: S
authors: Gouarin
authors: Joelle
authors: J
authors: Petitjean
authors: Jean-Jacques
authors: JJ
authors: Parienti
authors: Delphine
authors: D
authors: Nimal
authors: Jacques
authors: J
authors: Brouard
authors: François
authors: F
authors: Freymuth
authors: Astrid
authors: A
authors: Vabret
mesh_terms: D000258:Adenovirus Infections, Human / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D004267:DNA Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D005260:Female; D056661:Human bocavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D008875:Middle Aged; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012137:Respiratory System / Q000821:virology; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D004279:DNA, Viral