Application of Dried Whole Blood Collected on Filter Paper Disks to ELISA for the Detection of Sendai Virus and Mouse Hepatitis Virus Antibodies in Mice

1992 Experimental animals  
This study was undertaken to simplify the preparation procedures for test specimens by applying whole blood collected on filter paper disks. The results of ELISA obtained using specimens collected in this way for the detection of Sendai virus and mouse hepatitis virus antibodies in mice were comparable to those for ordinary ELISA using serum samples. KEY WORDS : MHV antibody, mouse, Sendai virus antibody, whole blood ELISA We developed ELISA kits for the detection of Sendai virus and mouse
more » ... irus and mouse hepatitis virus (MHV) antibodies in mice [2, 3] . Sera are used as the test specimens in these kits. Some users have complained of the troublesomeness of collecting the appropriate amount of blood and centrifuging it to prepare serum samples. Furthermore, the necessity of keeping sera frozen or cooled presents some difficulties in the transportation of test materials. Therefore, we have attempted to simplify the test specimen preparation procedures. One hundred and nine mice of various strains and stocks reared in 12 conventional facilities were used in this study. For the collection of blood samples, filter paper disks 5mm in diameter (Advance Toyo, Thin) were employed so that approximately 10 p 1 of blood could be absorbed on each disk. Two disks for each mouse were dipped into the blood sample and dried at room temperature. The disks with whole blood were kept in refrigerators for 48 to 69 days until testing except when left at room temperature during 2-3 days' transportation from the mouse rearing facilities to our laboratory. The two serum samples were collected simultaneously from each mouse. ELISA kits, MONILISA' HVJ (Sendai virus) and MHV (Wakamoto Pharmaceutical Co. Ltd., Tokyo), were used in this study. The whole blood paper disks collected in duplicate were transferred to the wells of the antigen plates, one for Sendai virus untibody detection and the other for MHV antibody detection, then 200p1 of PBS-containing 0.1% BSA was added to each well and the plates were left for 1 hour at 37t to release the serum into the buffer solution. After incubation, the paper disks were removed. These materials, as well as the . simultaneously collected serum samples diluted 40-fold, were submitted to ELISA. ELISA was performed according to the manufacturer's directions. After the reaction, OD values were evaluated using a spectrophotometer . OD values of 0.3 or greater were regarded as positive. The results shown in the table indicate that Sendai virus antibody detection in the two corresponding samples (whole blood and serum) agreed, with a positivity of 4.6% (5/ 109) in each.The correlation coefficient of the OD values was 0.97 (y =0.003+0.926x; y : sera, x : whole blood). Out of the 109 specimens, MHV antibody was detected in 23 (21.1%) for the dried whole blood, whereas it was detected in 29 (26.6%) for the serum samples. The OD
doi:10.1538/expanim1978.41.3_389 fatcat:rtugo4lohfdkfgmhbt2waxwamy