APHL 2022 POSTER ABSTRACTS
Whole genome sequencing ( WGS ) was completed on S . marcescens isolates from a subset of environmental samples and available patient specimens . Results : Of 152 environmental samples analyzed , 24 ( 15.8 %) were positive for S . marcescens , including a used needle / syringe surrendered by an inmate , used nasal spray bottle containing methamphetamine , inmate hand rinsate , unmaintained disinfection dilution machine , bottles , mop bucket and trash cans used for diluting , storing and dispensing CB64 , used bottles containing drinking water , floor surfaces and scrubbies . S . marcescens was not detected in water samples , rinsates from sharps containers with used needles , showers , walls and unopened bottles of CB64 concentrate . WGS identified a cluster of genetically similar S . marcescens strains from clinical specimens from five case-patients , the used needle and the nasal spray bottle ; strains from other patients and environmental sources were highly diverse and not related to strains in this cluster . A lab experiment demonstrated persistence and growth of S . marcescens inoculated into CB64 diluted at 1:1000 for up to seven days . S . marcescens failed to survive and grow in concentrated and properly diluted ( 1:64 ) CB64 .
Conclusions : An unusual outbreak of invasive S . marcescens occurred at a prison in 2020 – 21 . Epidemiologic investigation suggested that the pathogen was likely introduced primarily through IDU . Lab investigation suggested widespread contamination ; notably , improper preparation , storage and use of CB64 likely reduced its activity against S . marcescens leading to persistence and further spread in the prison environment .
Presenter : Donna Ferguson , Monterey County Public Health Laboratory , fergusond @ co . monterey . ca . us
Rapid , Economical qPCR testing for Detection of Waterborne Pathogens Onsite and in the Laboratory
W Ulloa , K Smith and B . Guzman , Nephros
Every year , there are roughly 7.2 million infections , 120,000 hospitalizations and 7,000 deaths caused by waterborne pathogens in the United States . With the increase in population size and aging infrastructure , these number are likely to rise . Healthcare and commercial facilities often have large , complex plumbing systems that promote growth of biofilms . Biofilms can harbor pathogenic microorganisms , such as Legionella pneumophila , which can lead to Legionnaires disease in immune-compromised patients . Routine testing for Legionella bacteria is primarily in compliance with ISO 11731:2017 , which adopts a culture-based method for environmental water testing . However , culturing Legionella is time consuming and labor intensive . In addition , not all Legionella species are culturable . Organizations such as the EPA and CDC have researched for other analytical methods that can provide quicker turnaround time for environmental waterborne pathogen testing . Quantitative polymerase chain reaction or qPCR is the primary molecular method for waterborne pathogen testing . It is cost effective and can provide results in a relatively short amount of time . Real -time PCR is capable of positive detection of viable but non-culturable ( VBNCs ) pathogens by targeting the specific genetic sequence of the organism of interest . In addition , with the use of multiplexing in qPCR , it introduces the ability to detect multiple waterborne pathogens in one single run . This can be a great screening tool for routine testing . At Nephros , we use a rapid waterborne pathogen detection method that can be deployed onsite or in a laboratory . Overall workflow consists of filter concentrating one liter of water using a 5-nanometer hollow fiber membrane to capture microbes ranging in size from protozoans to viruses . After our 1-step lysis step , we perform qPCR using our Legionella pneumophila and species Taqman primer / probe mix , which complies with ISO 12869:2019 . This method allows us to provide a negative or positive screening result with a turnaround time of less than two hours . Real-time PCR can be an essential tool used for negative screening of waterborne pathogens and adds an extra layer of surveillance for VBNC waterborne pathogens .
Presenter : Wesley Ulloa , Nephros , wesley . ulloa @ nephros . com
Feasibility of the Commercially Available ChromaCode ® Tick-Borne Pathogen HDPCR Assay for Use in Vector Screening of Field-Collected Ticks in New Jersey
J Occi 1 , D Woell 1 , M Cuadera 2 , K Cervantes 2 , M Pugliese 1 , J Hill 3 , R Siderits 1 ; 1 New Jersey Public Health and Environmental Laboratories , 2 New Jersey Department of Health Infectious & Zoonotic Disease Program , 3 Chromacode
Background : New Jersey is among the highest-incidence states for multiple tick-borne diseases , including Lyme disease , anaplasmosis , babesiosis and ehrlichiosis and is home to a variety of tick species capable of transmitting pathogens , including Ixodes scapularis , Amblyomma americanum , Dermacentor variabilis and the invasive Haemaphysalis longicornis . The ranges and abundance of some of these tick species are increasing , leading to increased disease risk . New Jersey already has a comprehensive mosquito-borne vector screening program encompassing all 21 counties and collaboration between the Departments of Health and Environmental Protection . Expanding this initiative to include surveillance of tick populations is expected to bring valuable data on vectors and human pathogens to better inform public health campaigns and evaluate the effectiveness of disease prevention strategies implemented in this high-incidence state .
Objectives : This study evaluated a commercially available tickborne pathogen ( TBP ) HDPCR assay from ChromaCode ® for use in screening relevant pathogens in field-collected ticks . This assay is a multiplex method capable of detecting nine unique tick-borne pathogens in a single reaction , however , it is designed primarily as a clinical assay . We evaluated the performance of the assay in four tick species to determine the feasibility of using the assay as a part of future statewide surveillance initiatives .
Results : The Chromacode ® assay initially proved to be robust in detecting pathogens in ticks . Preliminary accuracy and precision validation studies demonstrated the successful detection of pathogens in simulated samples , including co-infected I . scapularis . However , several important procedural limitations were observed , including the potential for misidentification with variations in concentrations of target DNA , and non-specificity of the Rickettsia spp . primer set to differentiate between pathogenic or commensal Rickettsia species . Furthermore , PHEL has recently observed dramatically reduced internal consistency with this assay , which is believed to be attributed to changes made when the assay updated
Infectious Diseases