British Standard Poliomyelitis Antisera Types 1, 2, and 3
Authors: F. T. Perkins, D. G. Evans
Journal: BMJ (1959)

POLIOMYELITIS IN SINGAPOREMErBxsf 1549 polioviruses; and (b) to avoid the potential neutralizing effect of monkey antisera against simian viruses.Undiluted culture fluid was mixed with an equal quantity of the typespecific rabbit antiserum, and after an incubation period of one hour at room temperature 0.2 ml. was distributed in each of 10 monkey-kidney-culture tubes.At the end of seven days the medium is changed to permit observation of the tubes for another seven days for the detection of minimal amounts of non-poliovirus cytopathogenic agents.It is essential that the fresh medium contain 0.1 ml. of the specific rabbit antiserum per tube to prevent the emergence of poliovirus.Complete neutralization was obtained, and no other cytopathogenic agent was found.However, if a cytopathogenic effect had appeared, it would be necessary to check whether it was poliovirus that escaped neutraliza- tion or some other agent.Tests for Residual Neurotropism in Monkeys.-Thefluid was tested in 35 cynomolgus monkeys by intracerebral (1 ml.) and intraspinal (0.1 ml.) inoculation of the undiluted culture fluid and of the dilutions indicated below.The diluted, as well as undiluted, culture fluids are tested as a check against a zone phenomenon, which has been observed with certain strains or with cultures containing mixtures of virus of differing degree of neurotropism.The intracerebrally inoculated monkeys must be observed for at least four weeks.and the intraspinally inoculated monkeys for at least three weeks.The results of these tests, published in J. Amer.med.Ass., 1957, 164, 1216, were as follows: Paralytic Effect in Monkeys Paralytic Effect in Monkeys Inoculated Intracerebrally with Inoculated Intraspinaily with 1 ml. of Indicated Dilution 0 1 ml. of Indicated Dilution Undiluted 10-1 Undiluted | 10-1 10-2 0/10 0/5 1/10 0/5 0/5Test for A bsenice of Neurotropism in Spinally Inoculated Chimpanzees.-Althoughthis need not be a standard test, fluid was tested in three chimpanzees devoid of spontaneously acquired antibody for any of the three types of poliovirus.Chimpanzees with spontaneously acquired antibody for any of the three types of poliovirus might also have some lesions in the spinal cord as a result of the natural infection, and therefore are not suitable for this type of test.Each chimpanzee was inoculated intraspinally in the region of the lumbar enlargement with 0.2 ml. of undiluted culture fluid (approximately 8 x 10' P.F.U.).They were observed for at least three weeks prior to sacrifice for histological examination.Neither paralysis nor lesions were observed in any of the nine chimpanzees used in these tests, and sections through the lumbar spinal cord indicated that the inoculum had been properly placed in the grey matter.Tests in Human Beings.-Aliquotswere fed to human beings between January and May, 1957, as described in J. Amer.med.Ass., 1957, 164, 1216.The dose consisted of 0.01 ml.(0.1 ml. of a 1:10 dilution) of culture fluid added to a teaspoonful of cherry syrup just before administration.No illness resulted and no cytopathogenic agents, other than Excretion of Virus and Development of Antibody by 5 Children Without Low-avidity or High-avidity Neutralizing Antibody for Any of the 3 Types of Poliovirus Following Feeding of 0.01 ml. of Culture Fluid (100,000 TCID) of the Type 2 Attenuated Strain at 3-Week Intervals Excretion of Indicated Antibody Developmentt Type of Virus for Indicated Type Age Child in 2 Low-avidity+ High-avidity Years High-avidity Cytopatho-pH Test