Supportive Data

The following data support the use of this assay for clinical testing.

Accuracy/Diagnostic Sensitivity and Specificity:

A study of 791 clinical specimens compared shell vial culture and this polymerase chain reaction (PCR) assay. Included in the study were 288 swab specimens (nasal, throat, rectal, skin), 125 eye specimens, 221 respiratory specimens (bronchial washings, sputa, bronchioalveolar lavage, tracheal secretions), 56 fresh tissue specimens, 72 stools, 5 urines, and 24 body fluids/other specimens. Specimens were inoculated into culture tubes and examined for cytopathic effects over a period of 14 days, and subsequently assayed with this LightCycler (LC) assay. Comparison of cell culture with LC PCR yielded the following: total specimens positive by LC PCR was 83 (stool=7; respiratory=8; tissue=3; swabs=29; eye specimens=30; miscellaneous= 1 and urine=4) and total specimens positive  by culture were 76 (stool=6; respiratory=7; tissue=3; respiratory swabs=28; eye specimens=29; and urine=2, miscellaneous = 1). Of the 83 total positive specimens, PCR detected approximately 10% more adenovirus infections compared with culture. This assay detected all 57 serotypes of adenovirus tested.

Supplemental Data (Spiking Studies):

To supplement the above data, 30 negative samples of various types (cerebrospinal fluid, ocular, respiratory, stool, urine, and plasma) were spiked with adenovirus positive control plasmid at the limit of detection (approximately 10 targets/microliter). The 30 spiked specimens were run in a blinded manner with 30 negative (non-spiked) specimens. One hundred percent of the spiked specimens were positive, and 100% of the non-spiked specimens were negative.

Analytical Sensitivity/Limit of Detection:

The lower limit of detection of this assay is 10 targets/microliter in specimen matrix.

Analytical Specificity:

No PCR signal was obtained from extracts of 150 bacterial, viral, parasitic, and fungal isolates that could cause similar disease or could be found as normal flora in sites normally tested for this organism.

Precision:

Interassay precision was 100% and intra-assay precision was 100%.

Reference Range:

The reference range for this assay is "Negative."

Reportable Range:

This is a qualitative assay and results are reported as negative or positive for targeted adenovirus DNA.