Lab Matters Fall 2023 | Page 41

APHL 2023 POSTER ABSTRACTS

Development and Implementation of a Canine Infectious Respiratory Disease Panel ( CIRD )

T . Spikes , Y . Mosley , H . Naikare , L . Whittington , D . Baughman , R . Cox , M . Coarsey , A . Kalantari ; Tifton Veterinary Diagnostic & Investigational Laboratory

Canines are the most popular companion animal in the world with nearly 50 million households having at least one dog in the United States alone . Due to their popularity , they are also one of the most often seen and treated animals at veterinary clinics . When these animals present with symptoms of an illness , the vet may order testing by collecting specimens from the animals that are then sent to a diagnostic lab . Tifton Veterinary Diagnostic and Investigational Lab intends to develop a diagnostic panel to test for the following canine infectious respiratory diseases : CAV , CHV , B . bronchiseptica , Mycoplasma canis / cynos , Strep . zooepidemicus , Influenza A , CRCoV , CDV and CPIV . This will be accomplished by employing stringent testing validation methods and utilizing Indical and Indimag nucleic acid extraction kits and real time PCR .

Presenter : Teddy Spikes , trs53790 @ uga . edu

Development of Novel Methods for Total Organic Fluorine Assessments in Environmental and Clinical Matrices

L . Duncan , M . Shafer ; Wisconsin State Laboratory of Hygiene

Background . The PFAS ( per- and polyfluoroalkyl substance ) crisis facing the world is one of most pressing environmental chemical pollution emergencies in modern times . PFAS are now ubiquitous in the environment due to their persistence , mobility and extensive history of use , having been used in numerous industrial and consumer applications , with little regulation , since the 1950s . Many PFAS resist chemical and biological degradation , can bioaccumulate , and have been associated with a diverse array of adverse health outcomes . Preventing PFAS contamination of environmental reservoirs and subsequent human exposures is challenging because there are numerous potential sources , many of which are very poorly characterized . In addition , only a small number of the > 5,000 known PFAS are typically quantified ( 20-40 at most ), which limits efforts to identify PFAS compounds that are characteristic of specific sources . Further complicating our understanding of environmental cycling and bioaccumulation is the fact that many important classes of PFAS and precursors undergo transformation in the environment and human body to PFAS compounds of specific enhanced toxicity concern . Approach . On-going PFAS research efforts at the University of Wisconsin , State Laboratory of Hygiene , are directed at providing a more complete characterization of PFAS in environmental and clinical matrices through the application of new and advanced methods . In parallel with the development of new methods , we are collecting major environmental sources / vectors of PFAS and subjecting them to detailed PFAS characterization in an effort to define source specific PFAS chemical fingerprints . Lexi Duncan , our APHL fellow is integrated into this exciting and dynamic research endeavor . Largely absent from the current toolbox of PFAS analytical approaches are methods for quantification of the TOTAL pool of PFAS . Methods for total PFAS will provide a more accurate assessment of the PFAS burden / contamination of the sample / environment , and in combination with targeted methods ( e . g . LC / MS / MS ) allow for assessment of the “ missing ” PFAS – i . e . what are we not quantifying with our targeted methods . The scale of the “ missing ” fraction can be used to prioritize additional discovery efforts . Here we report on our progress in developing methods for total PFAS using Combustion Ion Chromatography ( CIC ) to provide levels of total organic fluorine . Key CIC performance metrics for both aqueous and solid matrices will be presented and placed into context with targeted PFAS levels . Once the CIC methods are in place , we will communicate to a broad array of agencies , especially Public Health Labs , the potential ( and limitations ) of these new tools for total PFAS characterization .

Presenter : Martin Shafer , mmshafer @ wisc . edu

Enhancing Swine Diagnostics for the Detection and Differentiation of ASF and CSF from Other Septicemic Diseases : Development of Novel Multiplex High Throughput Taqman qPCR Panels and Point-of-care Isothermal Diagnostic Platforms

C . Hollis , L . Whittington , A . Kalantari , Y . Mosley , H . Naikare ; Tifton Veterinary Diagnostic & Investigational Laboratory

African swine fever ( ASF ), Classical swine fever ( CSF ), Pseudorabies virus ( PRV ), and Porcine Reproductive and Respiratory Syndrome ( PRRS ) are four viral diseases that can cause major problems for populations of swine . In addition to these viral diseases , other septicemic diseases like Erysipelas , Salmonellosis , Pasteurellosis , Streptococcosis , and Listeriosis can also cause herd fatality and significant economic loss . Because of the economic impact these diseases can have on the swine industry , screening for these diseases is an important part of surveillance and diagnosis . Diagnosis of these diseases becomes a challenge since the clinical manifestations from one disease to another can look quite similar . For this research there are two main objectives . First , to develop multiplex Taqman qPCR panels for the detection and differentiation of ASF , CSF , and PRRS from other economically significant septicemic diseases affecting swine . Currently , there are NAHLN protocols for detecting ASF and CSF in a PCR reaction with an internal control . We propose to have ASF , CSF , and PRRS , in addition to an internal control , in the same PCR reaction . This would result in cost savings , less consumption of reagents , and reduced turn-around-time by having three targets in one PCR reaction . In addition to this panel , two other panels will be created to screen for the septicemic diseases listed above . Development of these three panels will result in diagnostic solutions for multiple swine diseases . The second objective is to develop isothermal helicase dependent amplification ( HDA ) assays for potential point-of-care testing for ASF and CSF pathogens in resource limited settings . This technology uses an enzyme that can separate DNA strands at a constant temperature of 65 ° C . The use of a simple heat block can be used in the place of complex machinery for the reaction . We aim to provide proof of concept for the HDA assay and potentially incorporate this process into lateral flow assay strips . This will serve as a potential point-of-care testing option that can be performed barn side and allow rapid detection and diagnosis of ASF and CSF .

Presenter : Carol Hollis , carol . hollis @ uga . edu