Abstract
A sensitive gel-diffusion assay for determination of phospholipase A2 was developed. PLA2 standards, serum, faecal and pancreas homogenate samples with PLA2-activity were allowed to diffuse from wells into agar-gels containing lecithin-membranes. The turbidity cleared radially upon PLA2-activity. The diameters of the cleared zones showed a linear relationship with the log of the enzyme concentration. Serum samples resulted in some turbidity within the cleared zones. This interference originating from serum lipoproteins could be abolished by hydrophobic absorption. The gel-diffusion method was compared with two other methods for PLA2, titrimetric and radiometric techniques. Analysis on 37 human patients with acute pancreatitis showed close interrelationship between these methods. The phospholipase A2 activity in sera from man, the dog, the horse, the cow, the pig and the cat were almost equal, but much less than in the albino rat. No significant differences between PLA2 activities in pancreatic samples were obtained in different animal species. Of the faecal samples, the cow had the lowest PLA2 activity. Dogs suffering from pancreatic degenerative atrophy (PDA), had significantly reduced PLA2 activity both in their pancreas and faeces but not in serum.
Keywords: food poisoning, contamination, slaughter hygiene, microbial flora, public health
Sammanfattning
En känslig gel-diffusionsmetod för bestämning av fosfolipas A2 (PLA2) utvecklades. PLA2 standards, serum, feces- och pancreashomogenat med PLA2-aktivitet fick diffundera från fördjupningar i agargeler innehällande lecitinmembraner. Som en följd av PLA2-aktivitet skedde en cirkulär uppklaring av gelen. Diametrarna av de uppklarade zonerna stod i lineärt förhållande til logaritmen av enzymkoncentrationen. Serumprov resulterade i en vis grumlighet inom de uppklarade zonerna. Denna störning, som antagligen beror på lipoproteiner i serum, kunde avhjälpes med hydrofobisk absorption. Geldiffusionsmetoden jämfördes med två andra metoder för PLA2, en titrimetrisk och en radiometrisk teknik. Analys av 37 humanpatienter med akut pancreatit visade en nära samstämmighet med dessa metoder. Fosfolipas A2-aktiviteten i sera från människa, hund, hast, not, svin och katt var nästan lika, men mycket lägre an hos albinoråtta. Inga signifikanta skillnader mellan PLA2-aktiviteterna i pancreasprov från olika djurslag kunde konstateras. Bland de fecala provena hade nöt den lägsta PLA2-aktiviteten. Hundar med degenerativ pancreasatrofi (PDA), hade signifikant sänkt PLA2-aktivitet både i pancreas och feces, men inte i serum.
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