【摘要】：為建立黃熱疫苗病毒17D株的Vero細(xì)胞適應(yīng)株,將黃熱疫苗病毒在Vero細(xì)胞上培養(yǎng),通過蝕斑篩選,然后進(jìn)行適應(yīng)性傳代,并進(jìn)行穩(wěn)定性研究。 將雞胚黃熱疫苗病毒在Vero細(xì)胞上進(jìn)行了適應(yīng)性培養(yǎng),確定了傳代培養(yǎng)條件,MOI為0.1 PFU/cell,接種方式為分種,培養(yǎng)溫度為37℃。對2個批號的雞胚黃熱減毒活疫苗病毒進(jìn)行了2次蝕斑篩選,并連續(xù)傳代,獲得了YFV-Vero(5)-A1、YFV-Vero(5)-A2、YFV-Vero(5)-B1和YFV-Vero(5)-B2等4株Vero細(xì)胞適應(yīng)株病毒。經(jīng)過傳代后的穩(wěn)定性研究表明,4株Vero細(xì)胞適應(yīng)株病毒可被黃熱病毒(17D)猴免疫血清中和,證實(shí)確為黃熱17D株病毒。在傳代過程中不同代次的病毒滴度穩(wěn)定在6.9Lg PFU/ml以上,各代病毒蝕斑直徑大小沒有變化,高峰病變出現(xiàn)時間為5～7天。免疫原性研究結(jié)果表明,用BHK-21細(xì)胞和Vero細(xì)胞為基質(zhì)進(jìn)行中和試驗(yàn),家兔和小鼠的免疫血清均可中和3批不同批號的雞胚黃熱減毒活疫苗,中和抗體效價為1:1280～1:5120,顯示了4株Vero細(xì)胞適應(yīng)株病毒良好的免疫原性。 初步試驗(yàn)結(jié)果表明,已獲得的4株黃熱減毒活疫苗Vero細(xì)胞適應(yīng)株病毒,其病毒滴度高且穩(wěn)定,中和抗體效價高,免疫原性良好。以后需進(jìn)一步做猴體神經(jīng)毒力試驗(yàn)以證實(shí)其遺傳穩(wěn)定性,做病毒株的全序列測定以分析其基因穩(wěn)定性。
[Abstract]:In order to establish the Vero cell adaptation strain of yellow fever vaccine virus 17D strain, yellow fever vaccine virus was cultured on Vero cells, screened by plaque, then passaged adaptively, and the stability of the virus was studied. Chicken embryo yellow fever vaccine virus was cultured adaptively on Vero cells, and the subculture conditions were determined. MOI was 0.1 PFU/cell, and the culture temperature was 37 鈩，

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