Assay of prick test inoculum volume. I. Use and reliability of a gamma camera-based method

Introduction: In dermatology and allergy there are clinical research circumstances where very small amounts of substances introduced into the skin have to be measured "in vivo." An example is the assay of reagents injected by prick test. As injected volumes are very small, it is necessary to use indicators that can be measured at very low concentrations. In in vitro studies, gamma-emitting radioisotopes have been shown suitable for use as the indicators. In in vivo studies, except for instruments devised for specific research requirements, the measurement of small sources is taken with a common gamma camera. Objective: The purpose of the present study is to evaluate the experimental reliability of a gamma camera-based method to measure microvolumes labeled with radioisotopes and its suitable application in vive studies. Methods: Using a solution of 99m Tc-pertechnetate, we prepared, with precision pipettes, some sets of scalar volumes ranging from 1 micro to 200 picoliters, which correspond to activities between some micros and some hundreds of picocuries. The volumes were measured with a gamma camera both with and without a collimator. The overall reliability of the method under different experimental conditions was evaluated for sensitivity, precision, and accuracy. Last, a blind measurement was taken as a final check on the overall reliability of the method. Results: The volume-activity correlation appeared to be linear, with a Spearman coefficient higher than 0.99. The correlation straight lines of the measurements taken with and without a collimator proved that, in both cases, the linearity of the system did not change. The method showed a high degree of precision and accuracy. The maximum variation coefficient never exceeded 1.5% and the standard error 2%. The sampling error of the measured volumes was less than 8% in all the sets: up to 7% was due to the manual operations and to the technical characteristics of the micropipettes. The gamma camera measurement error ranged from 1% to 3%. The blind tests experimentally confirmed the overall reliability of the method. Conclusions: The method we studied proved highly reliable and inexpensive. Measurement errors are almost exclusively due to sampling errors. The gamma camera is a device any nuclear medicine department is equipped with, and a solution of 99m Tc-pertechnetate is readily available.