[Examination of the sample centrifugation time for emergency coagulation test].

The rapidity of coagulation testing is important for use as appropriate substitution therapy in patients with, or at risk of critical bleeding requiring massive transfusion. Whereas the ordinary method of coagulation testing is known to be slow, in a critically haemorrhaging patient, a rapid turnaround time of coagulation testing becomes indispensable. To find out if coagulation test results will be affected by a shortened centrifugation time, we measured PT (prothrombin time), APTT (activated partial thromboplastin time), FIB (fibrinogen) and PLT (platelet) in plasma, using different centrifugation times (10 min, 5 min, 3 min), and analyzed the measurements. We found that, whereas centrifugation time significantly affected the PLT count in plasma (10 min; 5.17 +/- 3.71 x 10(3)/microl, 5min; 28. +/- 26.9 x 10(3)/microl, 3min; 63.7 x 10(3)/microl), PT(10min; 14.6 +/- 5.76 sec, 5min; 14.7 +/- 5.84 sec, 3min; 14.9 +/- 6.40 sec), APTT (10min; 36.4 +/- 15.9 sec, 5min; 36.8 +/- 16.5 sec, 3min; 34.7 +/- 11.4 sec) and FIB(10min; 361 +/- 134 mg/dl, 5min; 356 +/- 132 mg/dl, 3min; 356 +/- 125 mg/dl) were not affected. These data suggest that shortening centrifugation time will have no significant effect on the value of PT, APTT and FIB, in an emergency situation.

[Role of the clinical laboratory for patients with massive transfusion undergoing elective surgery].

The microvascular bleeding resulting from the dilutional coagulopathy can occur when patients with massive blood loss are treated by infusing a lot of crystalloids, colloids, and red blood cell concentrates. For the management of dilutional coagulopathy and the appropriate replacement therapy of with coagulation factors and platelets, we usually monitor the patient's course of with platelet count, conventional coagulation tests such as the prothrombin time, the activated partial prothrombin time, and the fibrinogen concentration. The central clinical laboratory has a responsibility for an accurate and quick report of these test results of patients with massive transfusion. Furthermore, use of point care testing is of clinical value to fulfill a clinical demand in case with dilutional coagulopathy.

Development of an improved rapid BACpro® protocol and a method for direct identification from blood-culture-positive bottles using matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been incorporated into pathogenic bacterial identification methods and has improved their rapidity. Various methods have been reported to directly identify bacteria with MALDI-TOF MS by pretreating culture medium in blood culture bottles. Rapid BACpro® (Nittobo Medical Co., Ltd.) is a pretreatment kit for effective collection of bacteria with cationic copolymers. However, the Rapid BACpro® pretreatment kit is adapted only for MALDI Biotyper (Bruker Daltonics K.K.), and there has been a desire to expand its use to VITEK MS (VMS; bioMerieux SA). We improved the protocol and made it possible to analyze with VMS. The culture medium bacteria collection method was changed to a method with centrifugation after hemolysis using saponin; the cationic copolymer concentration was changed to 30% of the original concentration; the sequence with which reagents were added was changed; and a change was made to an ethanol/formic acid extraction method. The improved protocol enhanced the identification performance. When VMS was used, the identification rate was 100% with control samples. With clinical samples, the identification agreement rate with the cell smear method was 96.3%. The improved protocol is effective in blood culture rapid identification, being both simpler and having an improved identification performance compared with the original.