The superior sensitivity and specificity associated with the use of molecular assays has greatly improved the field of infectious disease diagnostics by providing clinicians with results that are both accurate and rapidly obtained. Herein, we review molecularly based infectious disease diagnostic tests that are Food and Drug Administration approved or cleared and commercially available in the United States as of December 31, 2010. We describe specific assays and their performance, as stated in the Food and Drug Administration's Summary of Safety and Effectiveness Data or the Office of In Vitro Diagnostic Device Evaluation and Safety's decision summaries, product inserts, or peer-reviewed literature. We summarize indications for testing, limitations, and challenges related to implementation in a clinical laboratory setting for a wide variety of common pathogens. The information presented in this review will be particularly useful for laboratories that plan to implement or expand their molecular offerings in the near term.
We compared the performances of the Third Wave Technology Invader method and the Digene Hybrid Capture 2 assay to detect high-risk human papillomaviruses in 87 cervical brushing specimens submitted in Cytyc ThinPrep media. Two different methods for the extraction of DNA from squamous epithelial cells were also evaluated.The human papillomavirus (HPV) family consists of more than 100 types. Those shown to be causative agents of cervical cancer are referred to as "high-risk" strains. Current guidelines recommend the testing of all women with a cytological diagnosis of atypical squamous cells of undetermined significance (ASCUS) for infection by high-risk types of HPV (16,23). HPV testing has been found to have a higher sensitivity than standard or liquid-based Pap testing for detecting high-grade cervical intraepithelial neoplasia (1,(17)(18)(19)(20). Only the Digene Hybrid Capture 2 (HC2) assay (Digene, Gaithersburg, MD) has been approved by the FDA for the detection of high-risk strains of HPV as an ASCUS triage for referral to colposcopy, and, in conjunction with cervical cytology, as a primary screen for cervical cancer. The purpose of this study was to compare a laboratory-developed test utilizing analyte-specific reagents (ASRs) developed by Third Wave Technologies Inc. (TWT, Madison, WI) with the Digene HC2 assay for the detection of high-risk HPV DNA in cervical brushings. The TWT method used in this study, based on proprietary Invader technology, employed isothermal signal amplification to detect 13 high-risk HPV types, utilizing three probe pools (A5/A6 [types 51 and 56], A7 [types 18, 39, 45, 59, and 68], and A9 [types 16, 31, 33, 35, 52, and 58]) based on phylogenetic relatedness. The assay also incorporated an internal control for human alpha-actin to control for DNA quality and quantity in each reaction.The 87 specimens used in this study were cervical brushings collected in ThinPrep fluid (Cytyc Corp., Marlborough, MA) submitted to the cytology laboratory at CompuNet Clinical Laboratories for routine Pap testing. Those samples that received a cytological diagnosis of ASCUS were tested for highrisk HPV by using the Digene HC2 high-risk HPV test according to the manufacturer's instructions. Cells were collected from 2 ml of ThinPrep fluid by centrifugation at 14,000 ϫ g for 10 min and suspended in 0.4 ml of phosphate-buffered saline.DNA was extracted using the Qiagen BioRobot M48 and the MagAttract virus DNA minikit (Qiagen Inc., Valencia, CA) and eluted into 100 l of RNase-free distilled water. Duplicate samples were prepared from 2 ml of each cytology sample by using Gentra Systems Puregene spin columns (Qiagen Inc., Valencia, CA) by following the manufacturer's instructions. All extracted samples were stored at Ϫ20°C until tested. The extracted DNA was tested for the presence of high-risk HPV by a laboratory-developed assay utilizing Invader HPV ASRs. The TWT ASRs consisted of three separate oligonucleotide pools with 6-carboxyfluorescein (FAM)-labeled probes, one Redmond red (RED)-labeled oligonu...
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