Development of a PCR-and hybridization-based assay (PCR Adenovirus Consensus) for the detection and the species identification of adenoviruses in respiratory specimens.
BACKGROUND
Antigen detection assays and viral isolation techniques are routinely used to detect adenoviruses (Ad) associated with respiratory infections, and the value of the polymerase chain reaction (PCR) has recently been assessed.


OBJECTIVE
This paper describes a PCR-hybridization-immunoenzymatic assay (PCR Adenovirus consensus) used to detect Ad and identify Ad species in respiratory specimens.


RESULTS
On seven representative serotypes Ad 12, Ad 3, Ad 7, Ad 11, Ad 1, Ad 8, Ad 4, the mean genome equivalents per ml and the mean 50% infectious doses per ml were 10(6.3)and 10(4), respectively. Using 362 nasal aspirates from children, Ad were detected by immunofluorescence (IF) and culture in 97 cases (27%), by the PCR-Ad hexon method in 107 cases (29.5%) and by the PCR Adenovirus Consensus method in 113 cases (31.2%); 13 samples were found positive by both PCR and negative by the IF and culture methods; five samples were only positive according to the PCR Adenovirus Consensus) method. The sensitivity, specificity, predictive positive value and predictive negative value of the PCR Adenovirus Consensus method were 97.9%, 93.2%, 84%, 99.1%, respectively. The method identified the species (sp) from 91 positive amplicons: 1 Ad sp A, 44 Ad sp B, 42 Ad sp C, 3 Ad sp E, and 1 Ad sp F; 85 isolates were identified by IF or the neutralisation in culture, and 86 by a PCR-RE digestion method. The PCR Adenovirus Consensus detected six positive samples that were negative according to the IF and culture methods, and it identified the precise species of nine IF-positive and culture-negative nasal aspirates.


CONCLUSIONS
The PCR Adenovirus Consensus technique is more efficient than the classical IF or culture techniques for the detection of Ad in respiratory samples. An internal control is included to validate the screening results, and specific probes are used to identify the Ad species.
Vabret
Astrid
A
Laboratory of Human and Molecular Virology, University Hospital, Av. G. Clemenceau, 14033 Caen, France.
Gouarin
Stéphanie
S
Joannes
Martine
M
Barranger
Come
C
Petitjean
Joëlle
J
Corbet
Sandrine
S
Brouard
Jacques
J
Lafay
Françoise
F
Duhamel
Jean-François
JF
Guillois
Bernard
B
Freymuth
François
F
D000258:Adenovirus Infections, Human / Q000175:diagnosis* / Q000821:virology*; D000260:Adenoviruses, Human / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D002648:Child; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods / Q000706:statistics & numerical data; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011237:Predictive Value of Tests; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology*; D012680:Sensitivity and Specificity; D012703:Serotyping; D013045:Species Specificity