Development and performance evaluation of a streamlined method for nucleic acid purification, denaturation, and multiplex detection of \u3ci\u3eBluetongue virus\u3c/i\u3e and \u3ci\u3eEpizootic hemorrhagic disease virus\u3c/i\u3e

Bluetongue virus (BTV) and Epizootic hemorrhagic disease virus (EHDV) possess similar structural and molecular features, are transmitted by biting midges (genus Culicoides), and cause similar diseases in some susceptible ruminants. Generally, BTV causes subclinical disease in cattle, characterized by a prolonged viremia. EHDV-associated disease in cattle is less prominent; however, it has emerged as a major economic threat to the white-tailed deer (Odocoileus virginianus) industry in many areas of the United States. The recent emergence of multiple BTV and EHDV serotypes previously undetected in the United States demonstrates the need for robust detection of all known serotypes and differential diagnosis. For this purpose, a streamlined workflow consisting of an automated nucleic acid purification and denaturation method and a multiplex one-step reverse transcription quantitative polymerase chain reaction for the simultaneous detection of BTV serotypes 1–24 and EHDV serotypes 1–7 was developed using previously published BTV and EHDV assays. The denaturation of double-stranded (ds) BTV and EHDV RNA was incorporated into the automated nucleic acid purification process thus eliminating the commonly used separate step of dsRNA denaturation. The performance of this workflow was compared with the World Organization of Animal Health BTV reference laboratory (National Veterinary Services Laboratory, Ames, Iowa) workflow for BTV and EHDV detection, and high agreement was observed. Implementation of the workflow in routine diagnostic testing enables the detection of, and differentiation between, BTV and EHDV, and coinfections in bovine blood and cervine tissues, offering significant benefits in terms of differential disease diagnosis, herd health monitoring, and regulated testing

A Retrospective Study Establishing Pre-Pandemic Demographic Baselines for United States Source Plasma Donors

Background The COVID-19 pandemic highlighted plasma as a strategic resource with continually increasing demand. Demographic data for plasma donors are limited compared with blood donors. Increased information regarding pre-pandemic donor demographics may serve as a baseline for evaluation of post-pandemic practices. OBJECTIVES This study asked the question, “What were the demographics of source plasma donors in the US compared to the general population and how did these change over a five-year period preceding the pandemic?” STUDY DESIGN Donor demographic data were retrospectively analyzed for the years 2014, 2016 and 2018 from a network of US plasma centers. METHODS Routine demographic data obtained prior to source plasma collection from all plasma donors at Grifols US centers were retrospectively analyzed. Donor screening and eligibility requirements were standardized across all donor sites and met all donor eligibility requirements (Code of Federal Regulations: 21 CFR part 600). During each calendar year, only data from the first donation in that year were included with each cohort year. RESULTS This study included 1,303,049 unique donors. Donors were predominantly young adult males, although females increased from 37.4% to 41.6%. Caucasians constituted the highest proportion, followed by African American and Hispanic donors. Demographics were generally stable, but the 2018 cohort and the US population exhibited significantly different age, race/ethnicity, and sex profiles. Of 2014 donors, 9.0% returned in all three years studied (2014-2016-2018), with a higher return rate of 16.0% observed for donors returning in just two of the years (2014-2016). Multiyear donors were predominantly male and African American. CONCLUSION US plasma donor demographics over a five-year period (2014-2018) showed generally consistent characteristics but differed from the general US population. Multiyear donors were demographically distinct from single-year donors. These data serve as a snapshot of the US source plasma donor base prior to the COVID-19 pandemic

Monoclonal antibody against novel epitopes of foot-and-mouth disease virus protein 3ABC and uses thereof

This disclosure pertains to isolated antibodies or antigen binding fragments thereof that specifically bind to the 3ABC non-structural protein of Foot-and-Mouth Disease virus (FMDV), wherein the antibodies or antigen binding fragments thereof recognize the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 12. Accordingly, this disclosure also pertains to polypeptides having an amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 12. Monoclonal antibody Mab 40C8 is also provided. The current disclosure also pertains to methods of detecting FMDV infection in an animal (including assays differentiating infected animals from vaccinated animals (DIVA)) and kits for performing the detection methods. Competitive ELISA kits comprising the antibody or antigen binding fragment thereof and immunoassay plates coated with the polypeptide comprising the amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 and/or SEQ ID NO: 12 are also provided.U

Monoclonal antibody against novel epitopes of foot-and-mouth disease virus protein 3ABC and uses thereof

This disclosure pertains to isolated antibodies or antigen binding fragments thereof that specifically bind to the 3ABC non-structural protein of Foot-and-Mouth Disease virus (FMDV), wherein the antibodies or antigen binding fragments thereof recognize the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 12. Accordingly, this disclosure also pertains to polypeptides having an amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 12. Monoclonal antibody Mab 40C8 is also provided. The current disclosure also pertains to methods of detecting FMDV infection in an animal (including assays differentiating infected animals from vaccinated animals (DIVA)) and kits for performing the detection methods. Competitive ELISA kits comprising the antibody or antigen binding fragment thereof and immunoassay plates coated with the polypeptide comprising the amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 and/or SEQ ID NO: 12 are also provided.U

Monoclonal antibody against novel epitopes of foot-and-mouth disease virus protein 3ABC and uses thereof

This disclosure pertains to isolated antibodies or antigen binding fragments thereof that specifically bind to the 3ABC non-structural protein of Foot-and-Mouth Disease virus (FMDV), wherein the antibodies or antigen binding fragments thereof recognize the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 12. Accordingly, this disclosure also pertains to polypeptides having an amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 12. Monoclonal antibody Mab 40C8 is also provided. The current disclosure also pertains to methods of detecting FMDV infection in an animal (including assays differentiating infected animals from vaccinated animals (DIVA)) and kits for performing the detection methods. Competitive ELISA kits comprising the antibody or antigen binding fragment thereof and immunoassay plates coated with the polypeptide comprising the amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 and/or SEQ ID NO: 12 are also provided.U

Monoclonal antibody against novel epitopes of foot-and-mouth disease virus protein 3ABC and uses thereof

This disclosure pertains to isolated antibodies or antigen binding fragments thereof that specifically bind to the 3ABC non-structural protein of Foot-and-Mouth Disease virus (FMDV), wherein the antibodies or antigen binding fragments thereof recognize the amino acid sequence of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 or SEQ ID NO: 12. Accordingly, this disclosure also pertains to polypeptides having an amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 12. Monoclonal antibody Mab 40C8 is also provided. The current disclosure also pertains to methods of detecting FMDV infection in an animal (including assays differentiating infected animals from vaccinated animals (DIVA)) and kits for performing the detection methods. Competitive ELISA kits comprising the antibody or antigen binding fragment thereof and immunoassay plates coated with the polypeptide comprising the amino acid sequence selected from SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 and/or SEQ ID NO: 12 are also provided.U

DS2_JVDI_10.1177_1040638718767943 – Supplemental material for Improvements in <i>Tritrichomonas foetus</i> molecular testing

<p>Supplemental material, DS2_JVDI_10.1177_1040638718767943 for Improvements in <i>Tritrichomonas foetus</i> molecular testing by Carly C. Ginter Summarell, Thomas B. Hairgrove, Megan E. Schroeder, Robert Conley, Mangkey A. Bounpheng in Journal of Veterinary Diagnostic Investigation</p

DS1_JVDI_10.1177_1040638718767943 – Supplemental material for Improvements in <i>Tritrichomonas foetus</i> molecular testing

<p>Supplemental material, DS1_JVDI_10.1177_1040638718767943 for Improvements in <i>Tritrichomonas foetus</i> molecular testing by Carly C. Ginter Summarell, Thomas B. Hairgrove, Megan E. Schroeder, Robert Conley, Mangkey A. Bounpheng in Journal of Veterinary Diagnostic Investigation</p

DS2_JVDI_10.1177_1040638718767943 – Supplemental material for Improvements in <i>Tritrichomonas foetus</i> molecular testing

<p>Supplemental material, DS2_JVDI_10.1177_1040638718767943 for Improvements in <i>Tritrichomonas foetus</i> molecular testing by Carly C. Ginter Summarell, Thomas B. Hairgrove, Megan E. Schroeder, Robert Conley, Mangkey A. Bounpheng in Journal of Veterinary Diagnostic Investigation</p

Development and performance evaluation of calf diarrhea pathogen nucleic acid purification and detection workflow

Calf diarrhea (scours) is a primary cause of illness and death in young calves. Significant economic losses associated with this disease include morbidity, mortality, and direct cost of treatment. Multiple pathogens are responsible for infectious diarrhea, including, but not limited to, Bovine coronavirus (BCV), bovine Rotavirus A (BRV), and Cryptosporidium spp. Identification and isolation of carrier calves are essential for disease management. Texas Veterinary Medical Diagnostic Laboratory current methods for calf diarrhea pathogen identification include electron microscopy (EM) for BCV and BRV and a direct fluorescent antibody test (DFAT) for organism detection of Cryptosporidium spp. A workflow was developed consisting of an optimized fecal nucleic acid purification and multiplex reverse transcription quantitative polymerase chain reaction (RT-qPCR) for single tube concurrent detection of BCV, BRV, and Cryptosporidium spp., and an internal control to monitor nucleic acid purification efficacy and PCR reagent functionality. In "spike-in" experiments using serial dilutions of each pathogen, the analytical sensitivity was determined to be <10 TCID50/ml for BCV and BRV, and <20 oocysts for Cryptosporidium spp. Analytical specificity was confirmed using Canine and Feline coronavirus, Giardia spp., and noninfected bovine purified nucleic acid. Diagnostic sensitivity was ≥= 98% for all pathogens when compared with respective traditional methods. The results demonstrate that the newly developed assay can purify and subsequently detect BCV, BRV, and Cryptosporidium spp. concurrently in a single PCR, enabling simplified and streamlined calf diarrhea pathogen identification.Keywords: Coronavirus, Cryptosporidium spp., Rotavirus, Reverse transcription quantitative polymerase chain reactio