Springfield - The Springfield laboratory is the primary site for the PHLS program. This facility is located on the Southern Illinois University School of Medicine campus at 825 N. Rutledge St. Also on campus are laboratory facilities for the Illinois Environmental Protection Agency, the Illinois State Police, and multiple SIU academic researchers. Courses are attended by either via video conference with SIU Carbondale, or on site with PHLS faculty. Blood Lead and Serology Syphilis - Screening blood for syphilis is done using the Rapid Plasma Reagin (RPR) agglutination card test. If positive, a Fluorescent Treponema Antibody (FTA) test is done for confirmation. As the name suggests a fluorescent dye is use to look for antibodies to the treponema which causes syphilis. These antibodies are visualized using a fluorescent microscope. Human Immunodeficiency Virus (HIV) - Screening for HIV is done using enzyme-linked immunosorbent assay (ELISA) test for the presence of antibodies to the HIV virus in blood or oral fluid. This is done using micro titer plates impregnated with HIV proteins. If the ELISA test is positive, a confirmatory Western Blot test is done. This detects the presence of antibodies to specific HIV proteins which are made visible by coloration. Clinical Chemistry - Analyze blood samples for blood lead from children 6 months - 6 years. Lead in blood is measured by Graphic Furnace - Atomic Absorption Spectrometry. The quantity of lead is measured based on light absorbed by ground state atoms of lead at 283.2 nm from a hollow cathode lamp source. Technologies/skills utilized: RPR agglutination, fluorescent antibody detection and microscopic examination, sample preparation and safety, GF-AAS, controlled dilutions, quality control, Western Blot Environmental Microbiology Dairy Products - This laboratory supports the Grade A Milk Program through testing of milk products and is recognized as the state central lab for the testing of milk by the FDA. Samples range from raw commingled milk to various finished products and are tested for quality (Petrifilm Aerobic Count), post pasteurization contamination (High Sensitivity Coliform Count), proper pasteurization, inhibitor, and Salmonella. Drinking Water - This laboratory also supports the enforcement of the Safe Drinking Water Act and is inspected once every three years by the US Environmental Protection Agency. Samples from various water supplies are analyzed for total Coliform-forming bacteria and E. coli using the Colilert system. Water samples include drinking water as well as that from public beaches. Foodborne Outbreak Investigations - This laboratory provides testing in support for foodborne outbreaks. Suspect food items from an outbreak are received and analyzed for specific pathogens based upon the epidemiological evidence. Samples are analyzed utilizing a combination of classical and automated methods for pathogen identification and characterization. Technologies/skills utilized: bacteriological culturing, microscopic examination, membrane filtering, Vitek 2 for organism characterization, sample preparation, Tecra Plus for sample screening, incubation, selective media Molecular Diagnostics Bioterrorism Response - The section responds to bioterrorism events and suspicious substances (e.g. white powders) by utilizing real time - polymerase chain reaction (RT-PCR) screening of samples within 2-4 hours for a panel of bioterrorism agents. This is followed by confirmatory testing in another section. Agents include: Yersinia pestis (plague), Bacillus anthracis (anthrax) and Francisella tularensis (rabbit fever). Sample types include: whole blood, nasal pharyngeal swabs, throat swabs, cultures, stools, vomit, plasma, serum, cerebrospinal fluid, tissue, and urine. Infectious Disease and other Outbreaks - PCR is utilized extensively. Stools are examined for Noroviruses (common causes of gastroenteritis) and their identification to the strain level to determine if organisms from different patients are related. Herpes Simplex Virus I and II can be detected in cerebrospinal fluid within 24 hours. Other agents subject to analysis include: enterovirus, rabies virus, Saint Louis Encephalitis virus, LaCrosse Encephalitis virus, Western Equine Encephalitis virus and West Nile virus (WNV) Technologies/skills utilized: Roche LightCycler real time polymerase chain reaction (RT-PCR), ABI 7000, ABI Fast 7500, Qiagen MDX automated sample preparation, work in Biosafety Level 3 (BSL-3) environment, pulsed-field gel electrophoresis (PFGE), gene sequencing, DNA hybridization Clinical Microbiology Rabies Analysis - Involves the removal of specific brain tissues from specimens suspected of having developed a rabies infection. The test is performed by specific examination of removed brain parts by Direct Fluorescence Antibody (DFA) techniques using dark field microscopic examinations. Bioterrorism Response - Requires becoming familiar with Centers for Disease Control and Prevention (CDC) procedures for the rule out and identification of biological terrorism agents in environmental and clinical specimens. Samples are received and prepared in a Biosafety Level 3 (BSL-3) environment and undergo confirmatory analysis for such agents as Yersinia pestis (plague), Bacillus anthracis (anthrax) and Francisella tularensis (rabbit fever), ricin and staphylococcal enterotoxin B, and botulinum toxin. Enteric Pathogens - Involves the identification of such pathogens as Salmonella spp, Shigella, E. coli O157:H7, Campylobacter, Vibrio spp, Yersinia spp and Staphylococcus aureus in clinical specimens through isolation, biochemical reactivity and selective media growth. Parasitology - Involves the examination of fecal specimens for protozoan cysts, helminth eggs and larvae, and trophozoites. Examination by formalin/ethyl acetate concentration, permanent stain procedure, acid fast staining techniques for cryptosporidium and cyclospora, and direct immunofluorescence for the simultaneous detection of Cryptosporidium oocysts and Giardia cysts. Sexually Transmitted Diseases - Involves molecular testing for the simultaneous amplification and detection of target DNA using amplification primers and a fluorescent-labeled detector probes. Processed samples are added to priming microplate wells, which contain the amplification primers, fluorescent-labeled detector probe, and other reagents necessary for the amplification and identification of Chlamydia trachomatis and Neisseria gonorrhea. Technologies/skills utilized: work in Biosafety Level 3 (BSL-3) environment, time resolved fluorescence (TRF) for bioterrorism agents, DIG ELISA for botulinum toxin, animal autopsy, preparation of reagents, tissue impressions, microscopic examination, quality control procedures, bacterial culture, sample dilution and preparation techniques, safety, incubation, pipeting and dilution techniques, ProbTec automated analysis, selective media, serological serotyping, slide smear preparations, wet mount and oil immersion, direct immunofluorescence, light and dark field microscopic examinations. Chicago - Depending upon logistics and preferences, students may be assigned to the Chicago laboratory. This facility is located on the campus of the University of Illinois at Chicago at 2121 W. Taylor St. This location is shared with the University's School of Public Health, Environmental and Occupational Health department. Courses are attended via video conference to either SIU Carbondale or Springfield. Carbondale - Depending upon logistics and preferences, students may be assigned to the Carbondale laboratory. This facility is located on the campus of the Southern Illinois University at 1155 S. Oakland Ave. Courses are attended by either video conference with Springfield, or on site with SIU Carbondale faculty.