D'Alessio D, Williams S, Dick E C
Appl Microbiol. 1970 Aug;20(2):233-9. doi: 10.1128/am.20.2.233-239.1970.
The use of fluorescein-conjugated antiserum against respiratory syncytial (RS) and parainfluenza 1 and 3 viruses was compared with conventional techniques in the rapid detection of virus in tissue cultures inoculated with pharyngeal specimens known to contain these viruses. Twenty-three specimens were tested: 9 RS, 8 parainfluenza 1, and 6 parainfluenza 3. The fluorescent-antibody technique (FA) detected virus in 52% of the tissue cultures in 24 hr, and, by 72 hr, 22 of the 23 cultures were FA-positive whereas only 5 were positive by conventional techniques. Additionally, conjugated antisera were prepared against herpes simplex, influenza A(2), and adenovirus type 5. All conjugates stained only the homologous virus and were 100- to 10,000-fold more sensitive than conventional techniques in detecting descending dilutions of virus inocula by 24 hr. With the procedures described, several antisera could be conjugated and ready for use within 24 hr. Serum fractionation was by ammonium sulfate precipitation, and with the procedure outlined virtually complete recovery of the globulin fraction and elimination of all of the albumin were accomplished.
将荧光素偶联抗血清用于呼吸道合胞体(RS)病毒、副流感1型和3型病毒的检测,并与传统技术进行比较,以快速检测接种已知含有这些病毒的咽拭子标本的组织培养物中的病毒。共检测了23份标本:9份RS病毒标本、8份副流感1型病毒标本和6份副流感3型病毒标本。荧光抗体技术(FA)在24小时内检测出52%的组织培养物中的病毒,到72小时时,23份培养物中有22份FA检测呈阳性,而传统技术仅检测出5份阳性。此外,还制备了针对单纯疱疹病毒、甲型流感病毒(2型)和5型腺病毒的偶联抗血清。所有偶联物仅对同源病毒染色,在检测病毒接种物24小时内的连续稀释液时,其敏感性比传统技术高100至10000倍。按照所述程序,几种抗血清可在24小时内偶联并准备好使用。采用硫酸铵沉淀法进行血清分级分离,按照所述程序几乎可完全回收球蛋白组分并去除所有白蛋白。