Publications by authors named "Ruth Hall-Sedlak"

Objective: To perform a diagnostic assessment of a point-of-care veterinary multiuse platform integrated with a model comprised of deep-learning, convolutional neural network algorithms for evaluating canine/feline peripheral blood smears compared to board-certified clinical pathologists (CPs).

Methods: This study had a blinded, randomized, incomplete block design, and results were compared between CPs and algorithms. Blood smears from convenience samples from veterinary diagnostic reference laboratories from October to December 2021 were used.

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The Vetscan Imagyst system (Zoetis) is a novel, artificial intelligence-driven detection tool that can assist veterinarians in the identification of enteric parasites in dogs and cats. This system consists of a sample preparation device, an automated digital microscope scanner, and a deep-learning algorithm. The EasyScan One scanner (Motic) has had good diagnostic performance compared with manual examinations by experts; however, there are drawbacks when used in veterinary practices in which space for equipment is often limited.

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Background: Fecal examinations in pet cats and dogs are key components of routine veterinary practice; however, their accuracy is influenced by diagnostic methodologies and the experience level of personnel performing the tests. The VETSCAN IMAGYST system was developed to provide simpler and easier fecal examinations which are less influenced by examiners' skills. This system consists of three components: a sample preparation device, an automated microscope scanner, and analysis software.

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Article Synopsis
  • Fecal examinations are essential for pet wellness, but their accuracy can vary based on sample preparation and the expertise of the person analyzing the results.
  • The VETSCAN IMAGYST system automates fecal analysis using advanced scanning and deep learning software to identify parasite eggs more efficiently compared to traditional methods.
  • Results showed that the VETSCAN IMAGYST's algorithm performed similarly to trained parasitologists, indicating high diagnostic reliability and potential advantages in ease of use and accuracy.
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  • The study examines how HHV-6B DNA is commonly found in human samples, highlighting the challenges in diagnosing its reactivation versus latency.
  • Researchers used RNA sequencing to analyze HHV-6B gene expression in different types of samples from patients, revealing similarities and differences in gene activity, especially focusing on the U38 gene, which showed consistent presence across all samples tested.
  • A new test targeting the U38 gene was developed, proving effective in identifying active HHV-6B infections, suggesting it could be a valuable tool for diagnosing lytic infections in patients without plasma detection of the virus.
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  • HHV-6A and -6B are highly prevalent human herpesviruses that can integrate into chromosomes, leading to inherited conditions like iciHHV-6.
  • Researchers sequenced the genomes of 61 active HHV-6B infection isolates from different locations and identified 64 strains of iciHHV-6B, revealing geographic clustering and evidence of genetic recombination.
  • The study provides a revised genome annotation for HHV-6B, enhancing our understanding of its genetic diversity and potential implications for detection and control of this virus.
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  • Human herpesvirus 6 (HHV-6) can integrate into chromosomes and be inherited through gametes, which leads to its presence in nearly every cell of individuals who have it.
  • A study analyzed 4319 pairs of hematopoietic cell transplant (HCT) donors and recipients to understand the impact of inherited chromosomally integrated HHV-6 (iciHHV-6) on post-transplant outcomes.
  • The presence of iciHHV-6 was linked to a higher risk of acute graft-versus-host disease and increased cytomegalovirus viremia, but did not significantly impact other outcomes like chronic GVHD or overall mortality, suggesting the potential for using iciHHV-6 screening in donor selection and
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  • HHV-6A/B can integrate their genomes into human chromosomes' telomeres, but the mechanisms and factors involved are not well understood due to the lack of effective cell culture models.
  • The study explored integration efficiencies of HHV-6A/B in various human cell lines using short-term infection and long-term growth methods, finding integration in multiple tested cell lines regardless of their telomerase status.
  • The research demonstrates that certain clones can express viral genes without producing infectious viruses, indicating potential avenues for further exploring drug effects on viral gene expression and integration mechanisms.
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  • Human rhinoviruses (HRV) include over 150 genotypes and are significant respiratory pathogens, necessitating effective molecular detection methods for diagnosis and surveillance.
  • Comparing reverse transcription-quantitative PCR (RT-qPCR) and reverse transcription-digital PCR (RT-dPCR), this study finds that RT-dPCR is more accurate for quantifying HRV RNA, especially with consensus primers that work across multiple genotypes.
  • RT-dPCR's advantage lies in its stability against sequence mismatches in primer/probe binding regions, making it a promising approach for quantifying diverse viruses like HRV in future studies.
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  • Human herpesvirus 6 (HHV-6) can integrate into chromosomes and is inherited in about 1% of the population through germ cells, potentially affecting human health.
  • A group testing approach was implemented to identify individuals with inherited ciHHV-6 by screening 2496 cellular samples from hematopoietic cell transplant (HCT) pairs, using advanced molecular techniques.
  • This efficient testing method confirmed inherited ciHHV-6 in a small percentage of donors and recipients, paving the way for larger studies on the clinical significance of this condition.
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  • Inherited ciHHV-6 in hematopoietic cell transplant (HCT) donors or recipients complicates testing for HHV-6 reactivation, which happens in 30-50% of cases.
  • A new multiplex droplet digital PCR assay can differentiate between HHV-6 species (A or B) and detect inherited ciHHV-6.
  • Testing showed that 25% of HCT recipients with ciHHV-6A experienced reactivation of HHV-6B, highlighting the importance of accurate diagnostic testing for HHV-6 infection despite the presence of ciHHV-6.
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Genome editing by designer nucleases is a rapidly evolving technology utilized in a highly diverse set of research fields. Among all fields, the T7 endonuclease mismatch cleavage assay, or Surveyor assay, is the most commonly used tool to assess genomic editing by designer nucleases. This assay, while relatively easy to perform, provides only a semi-quantitative measure of mutation efficiency that lacks sensitivity and accuracy.

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  • Diagnosis of Roseolovirus infections requires careful patient selection, sample choice, and testing methods.
  • Recent advances like quantitative PCR can distinguish between HHV-6 species A and B, but the benefits of screening high-risk patients are still uncertain.
  • Emerging techniques, such as digital PCR and reverse transcription qPCR, offer improved specificity and accuracy for diagnosing active infections despite challenges in result interpretation.
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We demonstrate the development of a multiplex droplet digital PCR assay for human cytomegalovirus (CMV), human adenovirus species F, and an internal plasmid control that may be useful for PCR inhibition-prone clinical samples. This assay performs better on inhibition-prone stool samples than a quantitative PCR assay for CMV and is the first published clinical virology droplet digital PCR assay to incorporate an internal control.

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We identified 37 hematopoietic cell transplantation recipients with human herpesvirus 6 (HHV-6) central nervous system dysfunction and tested donor-recipient pairs for chromosomally integrated HHV-6 (ciHHV-6). One patient had ciHHV-6A with possible HHV-6A reactivation and encephalitis. There was no ciHHV-6 enrichment in this group, but larger studies are needed to determine if patients with ciHHV-6 are at increased risk for HHV-6-associated diseases or other complications.

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  • The study investigates how often HHV-6 DNA is found in the cerebrospinal fluid of patients after hematopoietic cell transplantation (HCT) and its association with brain dysfunction.
  • Through testing 124 patients, researchers found HHV-6 DNA in 51, with 37 meeting criteria for neurological problems linked to the virus.
  • The findings indicate that while HHV-6 presence is common in those with neurological symptoms, those without such symptoms also showed poor survival rates, raising questions about the need for further treatment and research on these cases.
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Human cytomegalovirus (CMV) has historically been the major infectious cause of morbidity and mortality among patients receiving hematopoietic cell or organ transplant. Standard care in a transplant setting involves frequent monitoring of CMV viral load over weeks to months to determine when antiviral treatment may be required. Quantitative PCR (qPCR) is the standard molecular diagnostic method for monitoring.

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  • Despite the availability of an effective vaccine, hepatitis B virus (HBV) remains a significant public health issue due to costly treatments and the persistence of the virus in the form of covalently closed circular DNA (cccDNA) in liver cells.
  • Researchers developed zinc finger nucleases (ZFNs) aimed at disrupting the HBV genome to prevent viral replication and reactivation.
  • The study found that HBV-specific ZFNs could efficiently inhibit HBV replication and showed high specificity with minimal off-target effects, indicating their potential as a new treatment option for HBV eradication.
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  • Digital PCR (dPCR) is an emerging technology that may surpass real-time PCR (qPCR) in clinical virology diagnostics due to its enhanced precision and reproducibility.
  • Several studies indicate that dPCR assays for viruses like human cytomegalovirus and HIV produce more reliable results compared to traditional qPCR, while maintaining sensitivity.
  • The article reviews existing literature on the performance of dPCR versus qPCR and discusses the potential future applications of dPCR in the field of clinical virology.
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Background: Human herpesvirus 6 (HHV-6) latently infects a majority of adults. In about 1% of the population HHV-6 exists in a chromosomally integrated form (ciHHV-6) that resides in every somatic and germ cell and can be transmitted through the germ line. Patients with ciHHV-6 have been misdiagnosed and unnecessarily treated for active HHV-6 infection, sometimes with important side effects, based on results from quantitative molecular HHV-6 tests.

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Human cytomegalovirus (CMV) is a significant contributor to morbidity and mortality in immunocompromised patients, particularly in the transplant setting. The availability of anti-CMV drugs has improved treatment, but drug resistance is an emerging problem. Here, we describe an improved, rapid, sequencing-based assay for the two genes in CMV where drug resistance occurs, the UL97 and UL54 genes.

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  • Digital PCR (dPCR) allows for precise measurement of DNA without needing a standard curve, unlike traditional qPCR.
  • dPCR works by splitting a single reaction into many separate ones, using Poisson statistics to determine the quantity of DNA based on positive and negative signals.
  • The rising popularity of dPCR platforms is opening new opportunities in clinical diagnostics, especially for detecting low viral loads and rare mutations.
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  • Silver toxicity presents a challenge to microorganisms, with some bacteria adapting to grow in high silver ion concentrations, which can be both harmful as pathogens and potentially useful for nanomaterial production.
  • Researchers have engineered a silver-tolerant strain of Escherichia coli using a silver-binding peptide, AgBP2, which enhances the bacterium's tolerance to silver through a successful fusion with the maltose-binding protein (MBP).
  • The study finds that bacteria secreting the MBP-AgBP2 fusion in their periplasm can survive in environments with silver nitrate, revealing the potential for using engineered microbes in various applications, including environmental remediation and the study of metal interactions in living organisms.
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