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1.  Development of Multiplex PCRs for Detection of Common Viral Pathogens and Agents of Congenital Infections 
Journal of Clinical Microbiology  2005;43(10):5102-5110.
Potential causes of congenital infection include Toxoplasma gondii and viruses such as cytomegalovirus (CMV), enterovirus, hepatitis C virus, herpes simplex virus types 1 and 2 (HSV-1 and -2), human herpesvirus types 6, 7, and 8, lymphocytic choriomeningitis virus, parvovirus, rubella virus, and varicella-zoster virus. Testing for each of these agents using nucleic acid tests is time consuming and the availability of clinical samples such as amniotic fluid or neonatal blood is often limited. The aim of this study was to develop multiplex PCRs (mPCRs) for detection of DNA and RNA agents in the investigation of congenital infection and an mPCR for the viruses most commonly requested in a diagnostic virology laboratory (CMV, Epstein-Barr virus, enterovirus, HSV-1, HSV-2, and varicella-zoster virus). The assays were assessed using known pathogen-positive tissues (cultures, placentae, plasma, and amniotic fluid) and limits of detection were determined for all the agents studied using serial dilutions of plasmid targets. Nested PCR was performed as the most sensitive assay currently available, and detection of the amplicons using hybridization to labeled probes and enzyme-linked immunosorbent assay detection was incorporated into three of the four assays. This allowed detection of 10 to 102 copies of each agent in the samples processed. In several patients, an unexpected infection was diagnosed, including a case of encephalitis where HSV was the initial clinical suspicion but CMV was detected. In the majority of these cases the alternative agent could be confirmed using reference culture, serology, or fluorescence methods and was of relevance to clinical care of the patient. The methods described here provide useful techniques for diagnosing congenital infections and a paradigm for assessment of new multiplex PCRs for use in the diagnostic laboratory.
PMCID: PMC1248455  PMID: 16207970
2.  Epidemiology of group C rotavirus infection in Western New York women of childbearing age. 
Journal of Clinical Microbiology  1997;35(2):486-488.
Umbilical cord serum samples (380), an average of 10 per month for 3 years (1990 to 1992), were tested by indirect immunofluorescence assay for group C rotavirus immunoglobulin G. Thirty percent were positive, suggesting that approximately one-third of women of childbearing age in western New York have experienced group C rotavirus infection.
PMCID: PMC229607  PMID: 9003623
3.  Microtube coagulase test for detection of coagulase-positive staphylococci. 
Journal of Clinical Microbiology  1982;15(5):848-851.
Studies were performed to determine the sensitivity and specificity of a new microtube method for the detection of coagulase production by Staphylococcus aureus. Rabbit plasma containing EDTA was added to and lyophilized in API microtubes. Two standard coagulase plasmas containing EDTA were used in the conventional macrotube test and served as a basis for comparison. No false-positive or false-negative reactions were encountered with the microtube system. With this system, 53% of the coagulase-positive strains tested were detected within 1 h after inoculation, 82% were detected after 2 h, 97% were detected after 3 h, and 99% were detected after 4 h. With the first conventional method, 45, 81, 96, and 98% of the positive strains were detected in 1, 2, 3, and 4 h, respectively, whereas with the second conventional method, only 6% of the positive strains were detected in 1 h, 24% in 2 h, 66% in 3 h, and 81% in 4 h. With the microtube method, 5 of the 139 coagulase-producing strains studied reverted to negative between 5 and 24 h after inoculation, whereas 9 reverted with the more rapid conventional method, and no reversions occurred with the second conventional method. All reversions involved strains which caused gelation of plasma within 1 h after inoculation. The data obtained showed that 99% of the coagulase-positive strains tested could be detected within 4 h by the microtube method. In addition, the microtube method offers a more convenient and economical format for the performance of the coagulase tube test.
PMCID: PMC272200  PMID: 6808014

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