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Further Simplification of the Serum Neutralization Test with Herpes Simplex Virus
Author(s) -
Hondo Ryo,
Yoshino Kamesaburo
Publication year - 1972
Publication title -
japanese journal of microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0021-5139
DOI - 10.1111/j.1348-0421.1972.tb00664.x
Subject(s) - neutralization , herpes simplex virus , serial dilution , titer , neutralizing antibody , antibody , virus , virology , incubation , incubation period , plaque reduction neutralization test , chemistry , biology , immunology , medicine , pathology , biochemistry , alternative medicine
A new, simple technique for the serum neutralization test with herpes simplex virus was devised. Thin discs made of solidified overlay medium containing about 2×10 6 PFU/ml of virus (VACS) were covered with 0.01 ml of graded serum dilutions or control saline and placed on chick embryo cells with the serum‐covered face down. A 90‐mm monolayer dish received 8 such VACSs and then fixing overlay. The highest dilution of the serum which reduced confluent plaques to no or a few scattered plaques was taken as the endpoint. Incubation of the serum‐covered VACSs at 37 C for 1 hr prior to inoculation could be omitted, since this procedure did not markedly increase the endpoint. A 2‐ to 4‐fold fluctuation of the virus concentration had little influence on the endpoint. The standard deviation of the endpoint obtained was within ±2‐fold. The sensitivity of the test was about one‐third that of the 50% plaque reduction method. Early sera containing predominantly complement‐requiring neutralizing antibody exhibited considerable titers when tested by the new method even in the absence of complement, the reason for which remains unknown.