Correlation between amount of virus with altered nucleotide sequence and the monkey test for acceptability of oral poliovirus vaccine.

Production of live attenuated oral poliomyelitis vaccine (OPV) requires rigorous neurovirulence safety testing of each vaccine lot, currently carried out in monkeys. It has been reported that a change from 472-U to 472-C in the type 3 OPV RNA is associated with an increased histologic lesion score produced upon intraspinal inoculation of the mutant virus in monkeys. We have developed a method, based on polymerase chain reaction, for measuring the relative abundance of these mutant sequences directly in vaccine preparations and used this method to evaluate the proportion of 472-C in 40 different lots of type 3 OPV. Six vaccine lots that had failed the intraspinal monkey neurovirulence test contained a higher proportion of 472-C than all other lots that had passed this test. OPV type 3 virus containing 472-C was rapidly selected during serial passages in African green monkey kidney cells that are used for manufacturing of the vaccine. We have also found that the wild-type poliovirus type 3 strain Leon/37, from which the vaccine strain was originally derived, contained a mixture of 472-U and 472-C sequences. No other mutations in OPV type 3 RNA have been detected by similar assays at position 2034, also associated with attenuation, or at several other positions reported to be altered in some vaccine preparations. Our results suggest that molecular diagnostics may provide a supplement or a potential alternative to animal testing of live attenuated vaccines.

A new technique to improve cortical-evoked potentials in spinal cord monitoring. A ratio method of analysis.

Systemic effects such as anesthesia, hypotension, hypothermia, and hypoxia affect the cortical evoked responses. We propose, that by sequential stimulation of the median and posterior tibial nerves, and the construction of a ratio from the value of their amplitudes, the systemic effects can be eliminated and thus improve the reliability of the cortical evoked responses. Two groups of scoliosis patients who underwent spinal surgery with instrumentation were analyzed retrospectively. Both groups had spinal cord monitoring using peripheral nerve stimulation and cortical recordings of the somatosensory-evoked response (SER). In Group I, 50 patients were analyzed for changes in posterior tibial nerve response before and after distraction. Wide variability in the response suggested this method to be less reliable in predicting spinal cord conduction deficits. Thirty-eight patients in Group 2 were analyzed using both the median and posterior tibial nerve amplitudes. A ratio of the posterior tibial to median nerve wave amplitude was constructed, thus eliminating any systemic variables. A critical value, alerting the surgeons to possible decreases in spinal cord conduction, was calculated by subtracting one standard deviation from the mean of the postdistraction ratios of the posterior tibial to median nerves (1.20-.633 = .567).