Broadly reactive real‐time reverse transcription‐polymerase chain reaction assay for the detection of human sapovirus genotypes

Oka, Takahiro; Iritani, Nobuhiro; Yamamoto, Seiji; Mori, Kohji; Ogawa, Tomoko; Tatsumi, Chika; Shibata, Shuzo; Harada, Seiya; Wu, Fang‐Tzy

doi:10.1002/jmv.25334

Cited by 14 publications

(13 citation statements)

References 32 publications

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“…The sapovirus-specific sequence of M13F-HuSaV-5159F was identical to that of 1245Rfwd [15] except for 1 nt (Fig. 2 and Table 1), and they shared the same complementary sequence in the reverse primers (HuSaV-R, and SaV1245R, respectively) used in real-time PCR assays for detection of human sapoviruses [5,11].…”

Section: Contrast Four Assays Including the Primer Combinations I) Smentioning

confidence: 94%

“…The target regions of HuSaV-F1, -F2, and -F3 recently designed as forward primers in a broadly reactive real-time PCR for human sapoviruses [11] are similar to those of SV-F13 and SV-F14 (Fig. 2), and the sapovirus-specific sequence of M13R-HuSaV5498R (5′-CCCCANCCNGCVHACAT-3′) (Table 1) has the same target as the SVR-DS5 (5′-CCC CAC CCKGCC [11] are shown in parentheses. The primer combinations and PCR product size(s) are indicated at the top and right side, respectively, of each gel image.…”

Section: Contrast Four Assays Including the Primer Combinations I) Smentioning

confidence: 99%

“…This would cause a bias towards the detected strains or cause several genotypes to be missed when used for surveillance. We recently reported an improved broad-range real-time RT-PCR assay that detected all 18 of the currently identified human sapovirus genotypes [11]. This assay can be used as a screening tool, but the amplicon size (approximately 100 bp) is too short for further genetic characterization by sequence analysis.…”

Section: Contrast Four Assays Including the Primer Combinations I) Smentioning

confidence: 99%

“…To confirm the reactivity against clinical specimens, random-hexamer-primed cDNA synthesized from stool suspensions positive for sapoviruses (GI.1-7, GII.1-5 and GII.7 and GII.8, GIV.1, and GV.1-2) as well as stool suspensions positive for noroviruses (GI, GII), astrovirus types I and IV, and rotavirus groups A and C and DNA purified from stool suspensions positive for adenovirus type 40/41 that had been used in a previous study and stored at −30°C [11], were tested.…”

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confidence: 99%

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Polymerase chain reaction primer sets for the detection of genetically diverse human sapoviruses

et al. 2020

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Sapoviruses are increasingly being recognized as pathogens associated with gastroenteritis in humans. Human sapoviruses are currently assigned to 18 genotypes (GI.1-7, GII.1-8, GIV.1, and GV.1-2) based on the sequence of the region encoding the major structural protein. In this study, we evaluated 11 polymerase chain reaction (PCR) assays using published and newly designed/modified primers and showed that four PCR assays with different primer combinations amplified all of the tested human sapovirus genotypes using either synthetic DNA or cDNA prepared from human sapovirus-positive fecal specimens. These assays can be used as improved broadly reactive screening tests or as tools for molecular characterization of human sapoviruses.

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Section: Contrast Four Assays Including the Primer Combinations I) Smentioning

confidence: 94%

Section: Contrast Four Assays Including the Primer Combinations I) Smentioning

confidence: 99%

Section: Contrast Four Assays Including the Primer Combinations I) Smentioning

confidence: 99%

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confidence: 99%

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Polymerase chain reaction primer sets for the detection of genetically diverse human sapoviruses

et al. 2020

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“…Norovirus was tested by real-time RT-PCR as described previously 27 . Sapovirus, human astrovirus, and human parechovirus were tested by multiplex real-time RT-PCR using a QuantiTect multiplex PCR kit (QIAGEN, Hilden, Germany) and primer/probe sets as described previously 28 with a modification of the sapovirus primers/probes 29 .…”

Section: Clinical Sample Collection As Part Of Our Contribution To Tmentioning

confidence: 99%

Novel human reovirus isolated from children and its long-term circulation with reassortments

Yamamoto

Motooka

Egawa

et al. 2020

Sci Rep

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Mammalian orthoreovirus (MRV), also known as reovirus, was discovered in the 1950s and became the first reported segmented double-stranded RNA virus. MRVs have since been found in a variety of animal species, including humans. However, reports on MRV infections are scarce due to the rarity of their symptomatic occurrence. In Japanese surveillance studies, MRVs have been detected as gastrointestinal pathogens since 1981, with a total of 135 records. In Osaka City, Japan, MRV was first isolated in 1994 from a child with meningitis, and then in 2005 and 2014 from children with gastroenteritis. Here, we conducted the first molecular characterization of human MRV isolates from Japan and identified a novel human reovirus strain belonging to MRV type 2, designated the MRV-2 Osaka strain. This strain, with all three isolates classified, is closely related to MRV-2 isolates from sewage in Taiwan and is relatively close to an MRV-2 isolate from a bat in China. Our data suggest that the MRV-2 Osaka strain, which has circulated amongst humans in Japan for at least two decades, has spread internationally. The mammalian orthoreovirus (MRV) species belongs to the genus Orthoreovirus of the family Reoviridae, and is characterized by 10 segments of double-stranded RNA (dsRNA) genome encapsulated in the viral particles with a diameter of 65 to 80 nm 1. The ten dsRNA segments consist of three large (L1-L3), three medium (M1-M3), and four small (S1-S4) size class segments. The S1 segment encodes an attachment protein that binds to host cell receptors, thereby determining cell and tissue tropism, as well as the virus serotype 1. Four virus strains isolated from children in the 1950s (type 1 Lang, type 2 Jones, type 3 Abney, and type 3 Dearing) are the most frequently used MRV-1-3 prototypes to date. The fourth serotype (MRV-4) was isolated from a mouse, but its infection in humans has never been reported 2. Due to the segmented nature of MRV genomes, they are capable of undergoing gene reassortment during co-infection in one cell, resulting in the emergence of reassortants containing mixed segments from two or more different parental strains. Since there has been no type-specific segment found, except for the S1 segment, it seems that segments can be exchanged between any types of MRV. MRV has a wide geographic distribution, and is considered to infect practically all mammals, including humans 1. Human infections with MRV are common in early childhood 3,4 , but are rarely symptomatic 1. When MRV produces symptoms in human, the most common manifestations are coryza, pharyngitis, cough 5 , as well as gastroenteritis 6. More severe cases, including neurological diseases and meningitis, have also been reported 6-10. However, MRV is rarely detected in humans, even during longitudinal studies 6,11 , although it is one of the most abundant viruses in environmental waters 11,12. Here, we present epidemiological information regarding MRV detections in Japan since 1981. We conducted the first molecular characterization of MRV isolates from chi...

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Effect of Hydroxyapatite Nanorods on the Fate of Human Adipose‐Derived Stem Cells Assessed In Situ at the Single Cell Level with a High‐Throughput, Real‐Time Microfluidic Chip

Hao

Wang³

et al. 2019

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The fate of stem cells at the single cell level with limited communication with other cells is still unknown due to the lack of an efficient tool for highly accurate molecular detection. Moreover, the conditional sensitivity of biological experiments requires a sufficient number of parallel experiments to support a conclusion. In this work, a microfluidic single cell chip is designed for use with a protein chip to investigate the effect of hydroxyapatite (HAp) on the osteogenic differentiation of human adipose‐derived stem cells (hADSCs) in situ at the single cell level. By successfully detecting secretory proteins in situ, it is found that the HAp nanorods enhance osteogenic differentiation at the single cell level. In the chip, the single cell seeding approach confirms the osteogenic differentiation of the hADSCs, which endocytoses HAp, by reducing the influence of the factors secreted by neighboring differentiating cells. Most importantly, more than 7000 microchambers provide a sufficient number of parallel experiments for statistical analysis, which ensure a high level of repeatability of the HAp nanorod‐induced osteogenic differentiation. The microfluidic chip comprising single cell culture microchambers with in situ detection capability is a promising tool for research on cell behavior or cell fate at the single cell level.

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Broadly reactive real‐time reverse transcription‐polymerase chain reaction assay for the detection of human sapovirus genotypes

Cited by 14 publications

References 32 publications

Polymerase chain reaction primer sets for the detection of genetically diverse human sapoviruses

Polymerase chain reaction primer sets for the detection of genetically diverse human sapoviruses

Novel human reovirus isolated from children and its long-term circulation with reassortments

Effect of Hydroxyapatite Nanorods on the Fate of Human Adipose‐Derived Stem Cells Assessed In Situ at the Single Cell Level with a High‐Throughput, Real‐Time Microfluidic Chip

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