Pseudotyped Viruses: A Useful Platform for Pre-Clinical Studies Conducted in a BSL-2 Laboratory Setting

Biomolecules, Год журнала: 2025, Номер 15(1), С. 135 - 135

Опубликована: Янв. 15, 2025

The study of pathogenic viruses has always posed significant biosafety challenges. In particular, the highly requires methods with low biological risk but relatively high sensitivity and convenience in detection. recent years, pseudoviruses, which consist a backbone one virus envelope proteins another virus, have become most widely used tools for exploring mechanisms binding to cells, membrane fusion viral entry, as well screening libraries antiviral substances, evaluating potential neutralizing monoclonal antibodies, developing neutralization tests, therapeutic platforms. During outbreak severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), pseudotyped virus-based assays played pivotal role advancing our understanding virus-cell interactions its disease pathogenesis. Such facilitated search agents accelerated epidemiological studies on post-infection post-vaccination humoral immunity. This review focuses use pseudoviruses model large-scale applications enveloped viruses.

Язык: Английский

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Serological Assays Reveal No Evidence of Natural SARS-CoV-2 Infection in US Cattle

Microorganisms, Год журнала: 2025, Номер 13(3), С. 600 - 600

Опубликована: Март 5, 2025

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) continues to pose a significant threat public health. Notably, SARS-CoV-2 demonstrates the capacity infect various non-human animal species, including both captive and free-living animals. Earlier experimental studies revealed low susceptibility of domestic cattle (Bos taurus) ancestral B.1 lineage; however, recent findings indicate greater permissiveness Delta variant. While some detected evidence infection in Italy, Germany, India, Nigeria, currently, there is no infections US cattle. We have investigated over 600 samples, pre-pandemic pandemic sera collected from Pennsylvania for presence antibodies. Since serological tests inherent problems false positives negatives, we conducted comprehensive assessment multiple assays. As are known positive serum used hyperimmune raised with SARS-CoV-2-spike receptor binding domain (RBD) as control test validation. found that pseudovirus neutralization assays luciferase reporter system can produce results, care must be taken interpret diagnosis using these natural or transmission among US. This study underscores importance robust evaluation when employing detection populations.

Язык: Английский

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SARS-CoV-2 serosurvey of healthy, privately owned cats presenting to a New York City animal hospital in the early phase of the COVID-19 pandemic (2020-2021)

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Фев. 15, 2024

Abstract SARS-CoV-2, the cause of ongoing COVID-19 pandemic, not only infects humans but is also known to infect various species, including domestic and wild animals. While many species have been identified as susceptible there are limited studies on prevalence SARS-CoV-2 in Both non-domestic cats now established be infection by SARS-CoV-2. serious disease may occur some instances, majority infections appear subclinical. Differing data for reported, highly context-dependent. Here, we report a retrospective serological survey presented an animal practice New York City, located close proximity large medical center that treated first wave patients U.S. Spring 2020. We sampled 79, mostly indoor, between June 2020 May 2021, early part which time community was under strict public health “lock-down”. Using sensitive specific fluorescent bead-based multiplex assay, found overall 13/79 (16%) serologically-positive animals study period; however, Fall had confirmed positive 44%. For seropositive cats, performed viral neutralization test with live additionally confirm presence antibodies. Of thirteen 7/13 (54%) were virus neutralization, two previously documented respiratory signs, high titers 1/1024 1/4096; no statistically significant association seropositivity or breed, sex age Follow up sampling showed maintained over time. In comparison, 51% feline coronavirus (FCoV), endemic 30% negative FCoV. demonstrate impact defined population during humans, suggest human-cat transmission substantial our group. Our provide new context events across species. Significance has broad tropism can wide range leading expansion reservoir. Expansion this reservoir result accumulation mutations within these potentially giving rise variants facilitating reverse zoonotic transmission. Domestic particularly noteworthy regard due their contact humans. Currently, very stages especially United States. This addresses gap investigating seroprevalence epicenter pandemic States pandemic. work underscores importance adopting One Health approach prevention conducting routine surveillance different

Язык: Английский

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Comparative analysis of serological assays and sero-surveillance for SARS-CoV-2 exposure in US cattle

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Апрель 4, 2024

Abstract Coronavirus disease-2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) continues to pose a significant threat public health globally. Notably, SARS-CoV-2 demonstrates unique capacity infect various non-human animal species, documented in captive and free-living animals. However, experimental studies revealed low susceptibility of domestic cattle ( Bos taurus ) ancestral B.1 lineage infection, with limited viral replication seroconversion. Despite the emergence variants potentially altered host tropism, recent findings indicate greater permissiveness Delta variant infection compared other variants, though seroconversion no clear evidence transmission. While some detected antibodies Italy Germany, there is natural from United States or elsewhere. Since serological tests have inherent problems false positives negatives, we conducted comprehensive assessment multiple assays on over 600 serum samples, including pre-pandemic pandemic sera. We found that pseudovirus neutralization luciferase reporter system can produce positive results, care must be taken interpret diagnosis using these assays. transmission among USA. Hence, it critical develop more reliable tailored accurately detect populations rigorously evaluate diagnostic tools. This study underscores importance robust evaluation when employing for detection populations.

Язык: Английский

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Evaluation of Fecal Sample Pooling for Real-Time RT-PCR Testing SARS-CoV-2 in Animals

Viruses, Год журнала: 2024, Номер 16(11), С. 1651 - 1651

Опубликована: Окт. 23, 2024

During the COVID-19 pandemic, veterinary diagnostic laboratories tested both human and animal samples needed to ensure that they could accurately perform large numbers of tests in a timely manner. Sample pooling, methodology used effectively for over 80 years as surveillance tool screening potentially infected individuals, was employed. Given its sensitivity, real-time polymerase chain reaction (PCR) is more suitable employing this strategy, compared other less sensitive testing methods. In study, we evaluated capability detecting SARS-CoV-2 5-sample 10-sample pools feces using reverse transcriptase (rRT-PCR) well determined level sensitivity. A blinded method test (BMT) by an independent laboratory conducted assess five-sample fecal pool. To complement detection capability, stability genome within PBS suspension measured under various time temperature conditions across 28-day period. Our results showed limit 12.8 6.4 copies 25 µL PCR, respectively. The pooling resulted cycle threshold (Ct) value loss 2.35 3.45, Ct values known positive individual samples, but consistent still achieved containing with original below 36 34, simulation clinical all be detected regardless number (1-3) each BMT demonstrated excellent sensitivity (100 copies/reaction) RNA even though matrix effect observed. Finally, our show remains stable wide range variations. Overall, findings provide solid data scale up capacity laboratories.

Язык: Английский

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Nationwide seroprevalence of SARS-CoV-2 Delta variant and five Omicron sublineages in companion cats and dogs in the USA: Insights into their role in COVID-19 epidemiology

Emerging Microbes & Infections, Год журнала: 2024, Номер 14(1)

Опубликована: Дек. 5, 2024

Understanding SARS-CoV-2 epidemiology in companion animals is critical for evaluating their role viral transmission and potential as sentinels human infections. This large-scale serosurvey analyzed serum samples from 706 cats 2,396 dogs collected across the USA 2023 using a surrogate virus neutralization test (sVNT) to detect antibodies. Overall, 5.7% of 4.7% tested positive antibodies, with younger (under 12 months) showing significantly lower seropositivity rates (p = 0.0048). Additionally, we 153 variant-specific antibody responses six sVNT kits targeting Delta variant five Omicron sublineages. Among cats, 67.5% showed antibodies Delta, positivity sublineages follows: BA.1 (62.5%), BA.2 (42.5%), BA.4/BA.5 (77.5%), XBB (52.5%), XBB.1.5 (45.0%). In dogs, 55.8% were sublineage (46.0%), (37.2%), (58.4%), (13.3%), (9.7%). Given close contact between humans, persistence against various variants sublineages, our findings suggest that seroprevalence may serve valuable tool tracking COVID-19 epidemiology.

Язык: Английский

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