Development of duplex real-time PCR for detection of two DNA respiratory viruses.
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).
Dina
Julia
J
Department of Virology, University Hospital of Caen, Avenue Georges Clemenceau, 14033 Caen Cedex, France. dina-j@chu-caen.fr
Nguyen
Emilie
E
Gouarin
Stephanie
S
Petitjean
Joelle
J
Parienti
Jean-Jacques
JJ
Nimal
Delphine
D
Brouard
Jacques
J
Freymuth
François
F
Vabret
Astrid
A
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
D004279:DNA, Viral