Direct identification of chlamydiae from clinical samples using a DNA microarray assay: a validation study.
While DNA microarrays have become a widely accepted tool for mRNA expression monitoring, their use in rapid diagnosis of bacterial and viral pathogens is only emerging. So far, insufficient sensitivity and high costs have been the major limiting factors preventing more widespread use of microarray platforms in direct testing of clinical samples. In the present study, a total of 339 samples, among them 293 clinical specimens from animals and humans, were examined by the ArrayTube (AT) DNA microarray assay to detect chlamydial DNA and identify the species of Chlamydia and Chlamydophila involved. Samples included nasal and conjunctival swabs, formalin-fixed, paraffin-embedded and fresh organ tissue, milk, feces and cell culture. Notably, the AT test was shown to detect mixed infections in clinical samples. The calculated median sensitivity of 0.81 over the entire panel of clinical samples was comparable to conventional 16S PCR, but slightly lower than real-time PCR and other PCR assays. However, when a panel of long-time stored swab samples was excluded from the calculation, the sensitivity was clearly higher (0.87) and equivalent to that of real-time PCR. Altogether, the data demonstrate the suitability of this DNA microarray assay for routine diagnosis.
Borel
Nicole
N
Institute of Veterinary Pathology, University of Zurich, Switzerland.
Kempf
Evelyne
E
Hotzel
Helmut
H
Schubert
Evelyn
E
Torgerson
Paul
P
Slickers
Peter
P
Ehricht
Ralf
R
Tasara
Taurai
T
Pospischil
Andreas
A
Sachse
Konrad
K
D000818:Animals; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D002689:Chlamydia / Q000235:genetics* / Q000302:isolation & purification*; D002690:Chlamydia Infections / Q000453:epidemiology / Q000382:microbiology*; D003228:Conjunctiva / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics*; D004196:Disease Outbreaks; D005557:Formaldehyde; D006801:Humans; D008892:Milk / Q000382:microbiology; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards; D016612:Paraffin Embedding; D011200:Poultry; D018401:Sample Size; D012680:Sensitivity and Specificity; D012756:Sheep; D016707:Tissue Fixation
D004269:DNA, Bacterial; D005557:Formaldehyde