Determination of Porcine Blood Creatine Kinase by Firefly Luciferase

Abstract

The use of the creatine kinase-luciferase (CK-L) coupled reaction for porcine blood CK determination was studied. Adenosine triphosphate produced by CK is reacted with firefly L light-producing system. The method permits determination of CK in blood as opposed to traditional methods which require plasma or serum. The luciferase procedure was applied to nine barrows each of purebred Pietrain and Yorkshire (Y)-Minnesota No. 1 (M)-Pietrain (P) breeding [Yx M(PX M)]. Pietrain pigs exhibited higher (P<.05) ear-blood CK after transportation at 83 kg live weight, but not after exercise at an earlier age (40 kg live weight). Pietrain pigs exceeded YXM(PXM) pigs for longissimus (P<.01) and trapezius (P<.05) muscle pH decline rates, longissimus area (P<.01), ham plus loin percentage (P<.01), and percentage of four lean cuts (P<.01). YXM(PXM) pigs exceeded Pietrains for longissimus subjective color, morphology and marbling (all P<.01)and backfat thickness (P<.05). Intraclass correlation coefficients between blood CK following transportation stress and longissimus pH (20 min postmortem) were significant (P<.05) for Pietrain (r=--.67) and Y• M(PX M) pigs (r=-.79). Highly significant (P<.01)correlation coefficients were noted between serum CK, as determined by the Rosalki procedure, and blood CK, as determined by CK-L for Pietrain (r=.93) and for Yx M(PX M) pigs (r=.99). (

Blood Creatine Kinase as a Predictor of the Porcine Stress Syndrome

“…The development of live-animal predictive tests for the porcine stress syndrome (PSS) and pale, soft, exudative (PSE) muscle has received much recent attention. Currently, the most actively studied diagnostic procedures are the use of halothane anesthesia (Eikelenboom and Minkema, 1974), immunological blood typing (Rasmussen and Christian, 1976) and creatine kinase (CK) in serum (Addis et aL, 1974;Beermann et al, 1975) or in whole blood by the luciferase method (Hwang et al, 1977;Elizondo et al, 1977). The present study was conducted to investigate the use of blood CK as a predictor of PSS (as revealed by halothane anesthesia), including the statistical accuracy of the procedure.…”

“…Blood samples, in the form of blood spots collected on sample cards, were obtained from the ear following exercise stress at 28 -+ 1 weeks of age. CK was determined by the method of Hwang et al (1977) using an unpurifled luciferase preparation (Antonik, 1977).…”

“…This study provided a comparison of the efficacy of the luciferase test for blood CK (Antonik, 1977;Elizondo et al, 1977;Hwang et al, 1977) as a predictor of PSS in breed groups exhibiting high, intermediate and low (zero) incidences of PSS. Figure 1 illustrates a diagram for the screening method used.…”

Implications of skeletal muscle creatine kinase to meat quality

“…However, CK assays that allow the analysis of unprocessed blood samples, such as the bioluminescent method proposed by Hwang et al (1977) and the immunochemical method developed by Tuomola et al (2002), seems more suitable for application in animal production and slaughterhouse environments.…”