Hawthorne J D, Albrecht P
J Clin Microbiol. 1981 Apr;13(4):730-7. doi: 10.1128/jcm.13.4.730-737.1981.
A sensitive plaque neutralization assay for parainfluenza virus types 1, 2, and 3 and respiratory syncytial virus was developed in Vero and MA 104 cell cultures. The tests were performed in semimicrotoiter trays containing 24 wells, 16 mm in diameter. Parainfluenza virus type 1 formed plaques in Vero and MA 104 cells only when trypsin was added to the overlay medium. Plaquing of parainfluenza virus type 1 was more sensitive and technically reproducible in MA 104 cells than in Vero cells. Parainfluenza virus types 2 and 3 and respiratory syncytial virus readily formed plaques in Vero cells. Plaques with all viruses were necrotic in character, except for plaques produced by parainfluenza virus type 3, which appeared red due to an increased uptake of neutral red by infected cells. Different conditions for plaquing of the four viruses had to be used to obtain plaques of suitable size. Antibody titers of commercially prepared guinea pig typing sera were 5- to 50-fold higher by the plaque neutralization test than by complement fixation. The addition of guinea pig immunoglobulin G antiglobulin to the serum-virus mixtures enhanced the conventional neutralization test 5- to 10-fold. The sensitivity and specificity of the plaque neutralization test was also determined with sera of marmosets experimentally infected with parainfluenza virus types 1 and 3. The generally low postinfection titers could be enhanced, on the average, 40-fold by using human immunoglobulin G antiglobulin in the neutralization test. A low degree of cross-reactivity was shown between parainfluenza virus types 1 and 3 both in the conventional neutralization test and in the anti-immunoglobulin enhanced neutralization test.
在Vero细胞和MA 104细胞培养物中建立了一种用于检测1、2和3型副流感病毒以及呼吸道合胞病毒的灵敏蚀斑中和试验。试验在含有24个直径为16毫米孔的半微量滴定板中进行。仅当在覆盖培养基中添加胰蛋白酶时，1型副流感病毒才能在Vero细胞和MA 104细胞中形成蚀斑。1型副流感病毒在MA 104细胞中的蚀斑形成比在Vero细胞中更灵敏且技术上更可重复。2型和3型副流感病毒以及呼吸道合胞病毒能在Vero细胞中轻易形成蚀斑。除3型副流感病毒产生的蚀斑因受感染细胞对中性红摄取增加而呈红色外，所有病毒产生的蚀斑均为坏死性。必须使用四种病毒蚀斑形成的不同条件来获得大小合适的蚀斑。通过蚀斑中和试验测得的市售豚鼠分型血清的抗体效价比补体结合试验高5至50倍。向血清 - 病毒混合物中添加豚鼠免疫球蛋白G抗球蛋白可使传统中和试验增强5至10倍。还使用实验感染1型和3型副流感病毒的狨猴血清测定了蚀斑中和试验的敏感性和特异性。在中和试验中使用人免疫球蛋白G抗球蛋白，平均可使感染后通常较低的效价提高40倍左右。在传统中和试验和抗免疫球蛋白增强中和试验中，1型和3型副流感病毒之间均显示出低度交叉反应。
