The use of a platelet nucleotide assay as a possible diagnostic test for malignant hyperthermia.
A bioassay, using high-performance liquid chromatography (HPLC) analysis of platelet adenosine nucleotides and hypoxanthine, was studied for its potential use as a test for MH susceptibility. A protocol for the assay was developed, based on the method outlined by Solomons and Masson. The HPLC procedure was a rapid, efficient, sensitive, and highly reproducible technique for measuring ATP, ADP, AMP, and hypoxanthine in platelets. Conditions of extraction and storage were critical for preventing degradation of the nucleotides. Extraction of nucleotides at icebath temperature was found necessary. Storage of platelet extract in PCA, even at -20 degrees C, showed loss of ATP and ADP; hence, neutralization with KOH was essential before storage. Contrary to the findings of Solomons et al., the present study demonstrated that neither ATP depletion nor per cent reduction in nucleotide ratios in platelets treated with halothane can be used as a definitive test for the diagnosis of MH susceptibility. The reason for this disagreement is unclear; however, differences in methods and altitude are implicated. It is possible that the platelet is not affected by malignant hyperthermia and thus cannot serve as a test system for the detection of the syndrome.
D000227:Adenine Nucleotides / Q000097:blood*; D000244:Adenosine Diphosphate / Q000097:blood; D000249:Adenosine Monophosphate / Q000097:blood; D000255:Adenosine Triphosphate / Q000097:blood; D001792:Blood Platelets / Q000032:analysis* / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D005069:Evaluation Studies as Topic; D005260:Female; D006221:Halothane; D006801:Humans; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000097:blood*; D066298:In Vitro Techniques; D008297:Male; D008305:Malignant Hyperthermia / Q000097:blood* / Q000175:diagnosis; D012306:Risk; D013048:Specimen Handling / Q000295:instrumentation
D000227:Adenine Nucleotides; D007042:Hypoxanthines; D019271:Hypoxanthine; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D006221:Halothane