Monitoring shedding of five genotypes of RotaTeq vaccine viruses by genotype-Specific real-Time reverse transcription-PCR assays

Yuki Higashimoto, Masaru Ihira, Yu Miyazaki, Ayumi Kuboshiki, Sayaka Yoshinaga, Hiroyuki Hiramatsu, Ryota Suzuki, Masafumi Miyata, Hiroki Miura, Satoshi Komoto, Jun Yukitake, Koki Taniguchi, Yoshiki Kawamura, Tetsushi Yoshikawa

研究成果: Article

2 引用 (Scopus)

抄録

RotaTeq (RV5) is a widely used live attenuated pentavalent rotavirus (RV) vaccine. Although fecal shedding of RV vaccine strains persists for long time periods, it is unclear how each vaccine strain replicates in intestinal tissue and is excreted in stool. To examine this issue, we established RV5 genotype-specific real-time reverse transcription-PCR (RT-PCR) assays. Five real-time RT-PCR assays were designed for the VP7 gene in genotypes G1, G2, G3, G4, and G6. All assays exhibited excellent linearity, and the detection limit was 1 infectious unit (IU)/reaction for G2, G4, and G6 and 10 IUs/reaction for G1 and G3. No cross-reactivity was observed among G genotypes. The inter- and intra-assay coefficients of variation were less than 3%. The assays were used to examine 129 stool samples collected from eight infants who received RV5. In cases 1 and 2, who received three rounds of vaccination, RV shedding decreased gradually with the number of vaccinations. G1 and G6 shedding appeared to be predominant in comparison to shedding of the other genotypes. Patterns of fecal shedding of the five genotypes of vaccine viruses differed between the eight vaccine recipients. RV5 genotype-specific real-time RT-PCR assays will be useful to study the molecular biology of RV5 replication in infants and experimental animals.

元の言語English
ジャーナルJournal of clinical microbiology
56
発行部数6
DOI
出版物ステータスPublished - 01-06-2018

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Reverse Transcription
Vaccines
Genotype
Viruses
Polymerase Chain Reaction
Rotavirus Vaccines
Vaccination
Rotavirus
Limit of Detection
RotaTeq
Molecular Biology
Genes

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

これを引用

Higashimoto, Yuki ; Ihira, Masaru ; Miyazaki, Yu ; Kuboshiki, Ayumi ; Yoshinaga, Sayaka ; Hiramatsu, Hiroyuki ; Suzuki, Ryota ; Miyata, Masafumi ; Miura, Hiroki ; Komoto, Satoshi ; Yukitake, Jun ; Taniguchi, Koki ; Kawamura, Yoshiki ; Yoshikawa, Tetsushi. / Monitoring shedding of five genotypes of RotaTeq vaccine viruses by genotype-Specific real-Time reverse transcription-PCR assays. :: Journal of clinical microbiology. 2018 ; 巻 56, 番号 6.
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abstract = "RotaTeq (RV5) is a widely used live attenuated pentavalent rotavirus (RV) vaccine. Although fecal shedding of RV vaccine strains persists for long time periods, it is unclear how each vaccine strain replicates in intestinal tissue and is excreted in stool. To examine this issue, we established RV5 genotype-specific real-time reverse transcription-PCR (RT-PCR) assays. Five real-time RT-PCR assays were designed for the VP7 gene in genotypes G1, G2, G3, G4, and G6. All assays exhibited excellent linearity, and the detection limit was 1 infectious unit (IU)/reaction for G2, G4, and G6 and 10 IUs/reaction for G1 and G3. No cross-reactivity was observed among G genotypes. The inter- and intra-assay coefficients of variation were less than 3{\%}. The assays were used to examine 129 stool samples collected from eight infants who received RV5. In cases 1 and 2, who received three rounds of vaccination, RV shedding decreased gradually with the number of vaccinations. G1 and G6 shedding appeared to be predominant in comparison to shedding of the other genotypes. Patterns of fecal shedding of the five genotypes of vaccine viruses differed between the eight vaccine recipients. RV5 genotype-specific real-time RT-PCR assays will be useful to study the molecular biology of RV5 replication in infants and experimental animals.",
author = "Yuki Higashimoto and Masaru Ihira and Yu Miyazaki and Ayumi Kuboshiki and Sayaka Yoshinaga and Hiroyuki Hiramatsu and Ryota Suzuki and Masafumi Miyata and Hiroki Miura and Satoshi Komoto and Jun Yukitake and Koki Taniguchi and Yoshiki Kawamura and Tetsushi Yoshikawa",
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Higashimoto, Y, Ihira, M, Miyazaki, Y, Kuboshiki, A, Yoshinaga, S, Hiramatsu, H, Suzuki, R, Miyata, M, Miura, H, Komoto, S, Yukitake, J, Taniguchi, K, Kawamura, Y & Yoshikawa, T 2018, 'Monitoring shedding of five genotypes of RotaTeq vaccine viruses by genotype-Specific real-Time reverse transcription-PCR assays', Journal of clinical microbiology, 巻. 56, 番号 6. https://doi.org/10.1128/JCM.00035-18

Monitoring shedding of five genotypes of RotaTeq vaccine viruses by genotype-Specific real-Time reverse transcription-PCR assays. / Higashimoto, Yuki; Ihira, Masaru; Miyazaki, Yu; Kuboshiki, Ayumi; Yoshinaga, Sayaka; Hiramatsu, Hiroyuki; Suzuki, Ryota; Miyata, Masafumi; Miura, Hiroki; Komoto, Satoshi; Yukitake, Jun; Taniguchi, Koki; Kawamura, Yoshiki; Yoshikawa, Tetsushi.

:: Journal of clinical microbiology, 巻 56, 番号 6, 01.06.2018.

研究成果: Article

TY - JOUR

T1 - Monitoring shedding of five genotypes of RotaTeq vaccine viruses by genotype-Specific real-Time reverse transcription-PCR assays

AU - Higashimoto, Yuki

AU - Ihira, Masaru

AU - Miyazaki, Yu

AU - Kuboshiki, Ayumi

AU - Yoshinaga, Sayaka

AU - Hiramatsu, Hiroyuki

AU - Suzuki, Ryota

AU - Miyata, Masafumi

AU - Miura, Hiroki

AU - Komoto, Satoshi

AU - Yukitake, Jun

AU - Taniguchi, Koki

AU - Kawamura, Yoshiki

AU - Yoshikawa, Tetsushi

PY - 2018/6/1

Y1 - 2018/6/1

N2 - RotaTeq (RV5) is a widely used live attenuated pentavalent rotavirus (RV) vaccine. Although fecal shedding of RV vaccine strains persists for long time periods, it is unclear how each vaccine strain replicates in intestinal tissue and is excreted in stool. To examine this issue, we established RV5 genotype-specific real-time reverse transcription-PCR (RT-PCR) assays. Five real-time RT-PCR assays were designed for the VP7 gene in genotypes G1, G2, G3, G4, and G6. All assays exhibited excellent linearity, and the detection limit was 1 infectious unit (IU)/reaction for G2, G4, and G6 and 10 IUs/reaction for G1 and G3. No cross-reactivity was observed among G genotypes. The inter- and intra-assay coefficients of variation were less than 3%. The assays were used to examine 129 stool samples collected from eight infants who received RV5. In cases 1 and 2, who received three rounds of vaccination, RV shedding decreased gradually with the number of vaccinations. G1 and G6 shedding appeared to be predominant in comparison to shedding of the other genotypes. Patterns of fecal shedding of the five genotypes of vaccine viruses differed between the eight vaccine recipients. RV5 genotype-specific real-time RT-PCR assays will be useful to study the molecular biology of RV5 replication in infants and experimental animals.

AB - RotaTeq (RV5) is a widely used live attenuated pentavalent rotavirus (RV) vaccine. Although fecal shedding of RV vaccine strains persists for long time periods, it is unclear how each vaccine strain replicates in intestinal tissue and is excreted in stool. To examine this issue, we established RV5 genotype-specific real-time reverse transcription-PCR (RT-PCR) assays. Five real-time RT-PCR assays were designed for the VP7 gene in genotypes G1, G2, G3, G4, and G6. All assays exhibited excellent linearity, and the detection limit was 1 infectious unit (IU)/reaction for G2, G4, and G6 and 10 IUs/reaction for G1 and G3. No cross-reactivity was observed among G genotypes. The inter- and intra-assay coefficients of variation were less than 3%. The assays were used to examine 129 stool samples collected from eight infants who received RV5. In cases 1 and 2, who received three rounds of vaccination, RV shedding decreased gradually with the number of vaccinations. G1 and G6 shedding appeared to be predominant in comparison to shedding of the other genotypes. Patterns of fecal shedding of the five genotypes of vaccine viruses differed between the eight vaccine recipients. RV5 genotype-specific real-time RT-PCR assays will be useful to study the molecular biology of RV5 replication in infants and experimental animals.

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