NE Regional AIHA Conference Legionella Analytical Developments. Dec 5, PDF Free Download

Legionella Species That Cause Disease L. pneumophila (Lp) causes about 80 to 90% of Legionnaires Disease in the US. 75% caused by Lp serotype 1 (Lp1). Under diagnosed. Urinary Antigen test positive after day 5. Only looks for Lp1 In Australia, 80% of LD cases are caused by L. longbeachae A hospital in NYC had an outbreak caused by L. micdadeii Another hospital in NYC had an outbreak caused by Lp serotype 6 Hospital in NC had an outbreak due to L. feeleii

Legionella Species That Cause Disease L. pneumophila, L. anisa, L. oakridgensis, L. bozemanii, L. birminghamensis, L, brunensis, L. cherri,. L. cincinnatiensis,. L. dumoffii, L. erythra, L. fairfieldensis, L. feelei, L. gormanii, L. gratiana, L. hackeliae, L. israelensis, L. jamestownensis. L. jordanis, L. lansingensis, L. longbeachae, L. maceachernii, L. micdadei, L. moravica, L. parisiensis, L. quinlivanii, L. rubrilucens, L. santicrucis, L. sainthelensis, L. spiritensis, L. steigerwaltii, L. tucsonensis. L. wadsworthii, L. worsliensis Veterans Administration requires individual serotyping of Lp or complete species identification

Proper Analysis Culture is Gold Standard in US. CDC/ISO Method - Culture methods-10 species and individual serotyping of L. pneumophila 1-14 Molecular Methods - PCR Broad screen for 50 Legionella species combined, +/-, no individual identification/quantification - DNA sequencing of isolates for strain ID compare clinical outbreak strains to env outbreak strains

Isolating Legionella from Environmental Samples Isolation - Buffered Charcoal Yeast Extract Agar (BCYE) - With and Without Antibiotics Presumptive Presence plating suspected isolates on BCYE With and Without Iron and L-cysteine Confirmation/Identification- different results depending on type of kit purchased - Direct Fluorescent Antibody Technique (DFA) - Latex Agglutination

L. bozmanii

Interpreting Lab Results Can t draw conclusions based on one sample or one sample set Non -detectable samples - Limited sample number or volume Testing unconcentrated samples Culturing wastewater or potting soil samples without heat pre-enrichment Failing to run positive and negative control samples on media used Collecting unrepresentative samples Improper collection or handling of samples Positive samples Relatively common Not uncommon to find more than one type of Legionella No agreement on concentrations that are deemed hazardous (infectious dose is unknown; likely variable from person to person)

Field Test Kits Not CDC Elite Proficient. CDC will not accept into the Legionella proficiency program No validation of test method or procedure No validation of media used in the kits Is manufacturing of the media standardized? Reproduceability of the testing results not studied sufficiently Limit of Quantitation is not sensitive enough

How to Choose a Lab CDC Elite Certified or UK HSE proficient. Goal is to use an independent, third party testing lab to ensure no conflict of interest Ask lab what method they use and what the limits of detection are different methods may mean different results Beware of Labs that offer risk level in their reports Beware of consultants or water treatment providers doing their own Legionella testing. Beware of using field test kits for Heterotrophic Plate Count and Legionella Do not rely on HPC, ph, and/or free residual chlorine testing in lieu of Legionella testing

Thank You! Questions? Diane Miskowski 856-303-2528 or www.legionellatesting.com

References Legionella Regulations from HSE, WHO, South Australia, New South Wales Legionnaires Disease, OSHA Technical Manual, USDOL-OSHA, Washington, DC Guidelines for the Prevention of Nosocomial Pneumonia, Morbidity and Mortality Weekly Reports, CDC, Atlanta, GA, January 3, 1997 Hospital Associated Legionellosis, State of New York Department of Health, July, 2005 Approaches to Prevention and Control of Legionella Infection, Allegheny County Health Department, Allegheny County, PA January, 1997

References Report of the Maryland Scientific Working Group to Study Legionella in Water Systems in Healthcare Institutions, MD Department of Health and Mental Hygiene, Baltimore, MD, June 14, 2000 Procedures of the Recovery of Legionella from the Environment, US Centers for Disease Control, January, 2005 Water Quality-Detection and Enumeration of Legionella, ISO 11731 1&2 Legionella Human Health Criteria Document, USEPA, November, 1999

Break 30 minutes

NE Regional AIHA Conference Legionella Analytical Developments. Dec 5, 2014