Very low likelihood that cultivated oysters are a vehicle for SARS-CoV-2: 2021–2022 seasonal survey at supermarkets in Kyoto, Japan

Yamazaki, Yasuko; Thongchankaew-Seo, Uraiwan; Yamazaki, Wataru

doi:10.1016/j.heliyon.2022.e10864

Cited by 6 publications

(9 citation statements)

References 23 publications

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“…The PEG precipitation method customarily includes an overnight 4°C incubation process for virion capture (Lowther et al ., 2017; Yamazaki et al ., 2022). However, the NIHS protocol (2010) states that the virion-PEG complex can be recovered from the oyster midgut gland immediately by centrifugation at 8,000 g for 20 min without an overnight incubation.…”

Section: Discussionmentioning

confidence: 99%

“…15 ml of SAP (Kyokuto) was added and incubated for 15 min at room temperature. Then, 13.2 ml of PEG solution (40% PEG-NaCl, see details in our previous publication, Yamazaki et al ., 2022) was added and thoroughly mixed by vortexing, followed immediately by a second centrifugation at 8,000 g for 20 min. The supernatant was carefully removed after the second centrifugation.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…The Polyethylene glycol (PEG) precipitation method has been used worldwide to enrich and detect norovirus and other viruses from oysters (Lowther et al ., 2019; National Institute of Health Sciences, Japan (NIHS) 2010; Yamazaki et al ., 2022). While this method can concentrate any virus, the presence of sample-derived inhibitors reduces concentration performance (Lowther et al ., 2019; Miura et al ., 2018; Yamazaki et al ., 2022). In our previous studies, we have shown that a combination of a very short, low centrifugation process (900 g , 1 min) and a normal centrifugation process (10,000-20,000 g , 5 min) as a pretreatment step for simple concentration detection of target bacteria in chicken cecal contents (Sabike et al ., 2016).…”

Section: Introductionmentioning

confidence: 99%

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Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva

Yamazaki,

Alonso,

Galay

et al. 2024

Preprint

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Background: Controlling novel coronavirus pandemic infection (COVID-19) is a global challenge, and highly sensitive testing is essential for effective control. The saliva is a promising sample for high-sensitivity testing because it is easier to collect than nasopharyngeal swab samples and allows large-volume testing. Results: We developed a simple SARS-CoV-2 concentration method from saliva samples that can be completed in less than 60 min. We performed a spike test using 12 ml of saliva samples obtained from healthy volunteer people, and the developed method performance was evaluated by comparison using a combination of automatic nucleic acid extraction followed by RT-qPCR detection. In saliva spike tests using a 10-fold dilution series of SARS-CoV-2, the developed method was consistently 100-fold more sensitive than the conventional method. Conclusions: The developed method can improve the sensitivity of the SARS-CoV-2 test using saliva and speed up and save labor in screening tests by pooling many samples. Furthermore, the developed method has the potential to contribute to the highly sensitive detection of various human and animal viral pathogens from the saliva and various clinical samples.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva

Yamazaki,

Alonso,

Galay

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Then, they were centrifuged at 15,557 g for 30 min, The supernatant of 24 ml, equivalent to 80% of the total liquid volume, was transferred to a new 50ml-sterile plastic tube. After adding 9.6ml of PEG-NaCl solution equivalent to 40% of the total liquid volume to the new 50 ml-sterile plastic tube (See details of the components of PEG-NaCl solution in our previous publication, Yamazaki et al ., 2022). The tubes were then vortexed and quickly centrifuged at 8,000 g for 20 min to precipitate the ASF virions.…”

Section: Methodsmentioning

confidence: 99%

Development of a simple and highly sensitive method for the concentration and detection of African swine fever virus in oral fluids and its feasibility assessment on raised pigs in Northern Vietnam

Mai Thi,

Tran Thi Huong,

Dong Van

et al. 2024

Preprint

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Background: Early detection and early slaughter through quarantine are essential to prevent the spread of the African swine fever virus (ASFV). Highly accurate testing is effective for early detection, but it is still difficult to establish a system, especially in the Global South. Pooled oral fluids tests have been used for simple pathogen monitoring, but compared to blood tests, the virus concentration in oral fluids is low, resulting in false negative and missing true positive cases. We collected oral fluids from raised pigs in northern Vietnam and attempted a highly sensitive ASFV survey using a newly developed virus concentration and detection method. Results: In a spike test, the developed method showed up to 100 times greater sensitivity than a reference method. To compare and evaluate the performance of the developed method, a total of 68 pooled oral fluid samples were collected, 63 from northern Vietnam and 5 from southern Japan. Using real-time PCR, 9/68 (13.2%) were positive by the reference method, and 23/68 (33.8%) by the developed method. Using real-time LAMP, 1/68 (1.5%) were positive by the reference method and 6/68 (8.8%) by the developed method. Conclusions: The developed method improved the sensitivity of ASFV detection from oral fluids and enabled early diagnosis of pigs before the onset of the disease. The developed method has the potential to enable simple and highly sensitive diagnosis of ASF, which is essential for its early diagnosis and effective surveillance.

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Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva

Yamazaki,

Alonso,

Galay

et al. 2024

Heliyon

View full text Add to dashboard Cite

Very low likelihood that cultivated oysters are a vehicle for SARS-CoV-2: 2021–2022 seasonal survey at supermarkets in Kyoto, Japan

Cited by 6 publications

References 23 publications

Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva

Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva

Development of a simple and highly sensitive method for the concentration and detection of African swine fever virus in oral fluids and its feasibility assessment on raised pigs in Northern Vietnam

Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva

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