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Evaluation of the SmartCycler II system for real-time detection of viruses and Chlamydia from ocular specimens
Regis P Kowalski1, Paul P Thompson, Paul R Kinchington
1Charles T. Campbell Ophthalmic Microbiology Laboratory, University of Pittsburgh Medical Eye Center, Ophthalmology and Visual Sciences Research Center, University of Pittsburgh School of Medicine, Pittsburgh, Pa. 15213, USA.
Objective:
To compare the SmartCycler II system (Cepheid, Sunnyvale, Calif) results with those of standard cell culture, to compare the SmartCycler II system results with those of a dedicated polymerase chain reaction facility, and to establish the SmartCycler II system as a polymerase chain reaction method for detecting viral and chlamydial DNA from ocular specimens.
Methods:
True-positive samples (test-positive specimens based on standard testing) and true-negative samples (test-negative specimens based on standard testing) were processed for polymerase chain reaction using the SmartCycler II system for adenovirus, herpes simplex virus type 1, varicella-zoster virus, and Chlamydia trachomatis. Sensitivity, specificity, positive predictive value, negative predictive value, and efficiency were based on the testing of true-positive and true-negative specimens.
Results:
The descriptive statistics for adenovirus, herpes simplex virus type 1, varicella-zoster virus, and C trachomatis were, respectively, as follows: sensitivity, 85%, 98%, 100%, and 94%; specificity, 98%, 100%, 100%, and 100%; positive predictive value, 98%, 100%, 100%, and 100%; negative predictive value, 85%, 91%, 100%, and 98%; and efficiency, 92%, 95%, 100%, and 99%. Test sensitivity for the SmartCycler II system was equivalent to that from a central molecular laboratory.
Conclusion:
The descriptive statistics of the SmartCycler II system obtained in a small laboratory were comparable to those of a central molecular laboratory for detecting viruses and Chlamydia species. Clinical Relevance Polymerase chain reaction has great potential in the routine diagnosis of ocular infections in any conventional laboratory.
