JP2006317329A - Detection method for microbial contamination of blood infusion preparation using inclination of concentration consumption curve of dissolved oxygen in blood infusion preparation as index - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a novel detection method of the microbial contamination of a blood infusion preparation using the inclination of the concentration consumption curve of dissolved oxygen in the blood infusion preparation. <P>SOLUTION: The microbial contamination of the blood infusion preparation can be detected before the blood infusion of the blood infusion preparation preserved over a long period of time by measuring the inclination of the concentration consumption curve of dissolved oxygen in the blood infusion preparation. The detection of the microbial contamination of the blood infusion preparation inclusive of the autoinfusion of blood using a dissolved oxygen concentration measuring method is useful. As a method capable of detecting the microbial contamination of the blood infusion preparation, a rapid and automatic low cost practical system is constituted. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention is a method for detecting the slope of a dissolved oxygen concentration consumption curve in a blood transfusion preparation as an indicator of microbial contamination of a blood transfusion preparation and the slope of a dissolved oxygen concentration consumption curve in a blood transfusion preparation as an indicator of microbial contamination of the blood transfusion preparation. It is about.

  Bacterial contamination with transfusion products poses a fatal risk for patients receiving blood transfusions. In Japan, the MAP preparation is stored for 5 days in a refrigerated state, and the platelet preparation is stored at a normal temperature of 20-24 ° C. for a period of 3 days. If it is contaminated with bacteria during this storage period, for example, if Yersinia enterocolitica that grows at 4 ° C is contaminated, it will cause bacteraemia to the recipient after blood transfusion, leading to death There is an accident. In addition, since platelet preparations are stored at room temperature, there are accidents in which many bacteria are similarly contaminated and cause serious infections. Therefore, early detection is necessary for bacterial contamination in transfusion products.

Nippon Pall Co., Ltd. (PALL, USA) took 4 mL from the blood product bag and left it at room temperature for 24 hours, and then mixed the medium tablet (tripticase soy) and complement inhibitor at 35 ° C for 24 hours. After culturing, an oxygen gas detector tube is inserted into the gas phase part of the bag, and a system that measures the decrease in oxygen concentration with the BDS Oxygen Analyzer is completed and commercialized (eBDS). However, dissolved oxygen in transfusion products is not measured. A small bag is prepared for one blood donor, blood is collected, and gaseous oxygen is measured in a sealed state. Equipment development was not in time when the US FDA approved. BioMerieux (Japan, France, USA) developed a system that collects 5 mL of a blood transfusion product under the trade name Bactalart, inoculates it into a liquid medium for enrichment, and detects production due to bacterial respiration with a highly sensitive CO ₂ detector. Approved by the FDA. Originally, a commercial product was used as a bacteria detection device for blood of patients with bacteriophage deficiency.

Daikin Industries, Ltd. has developed and commercialized DOX ^™ , a contamination bacteria detection device for foods. This time we will use this oxygen electrode device to measure the oxygen concentration consumption in the solution of transfusion products and detect bacterial contamination. In Non-Patent Document 1, the growth of bacteria in a medium is observed using oxygen consumption as an index, but there is no observation of the growth of bacteria in a blood transfusion preparation with the slope of a dissolved oxygen consumption curve.
Japanese Patent Application No. 2000-81765 Japanese Patent Application No. 2000-81766 Japanese Patent Application No. 2000-76393 Machida Katsuhiko, Symposium 1: Problems with drug-resistant bacteria, Trial of speeding up susceptibility testing, Journal of Japanese Clinical Microbiology, 9 (2) 178-183, 1999

  Problems to be solved are a method for measuring microbial contamination of a new blood transfusion product, a method for providing a microbial contamination index for a new blood transfusion product, and a method for detecting the same.

  The most important feature of the present invention is that the blood transfusion preparation is artificially inoculated with bacteria, continuously cultured, and the slope of the dissolved oxygen consumption curve can be found.

  According to the present invention, a dissolved oxygen concentration measurement method is used to collect a part 24 hours after production, which is the time of manufacture of a blood transfusion preparation or at the time of delivery to a facility where the preparation is used, and continuously measure the dissolved oxygen concentration after about 20 hours. Is detectable. After 48 hours of production, which is a typical standard time for blood transfusion, the state of microbial contamination of the transfusion product can be known in advance. As a result, there is a high possibility that an accident during blood transfusion can be avoided. A rapid, automated, low-cost practical system for detecting microbial contamination. Detection of microbial contamination of blood products, including autologous blood transfusion using dissolved oxygen concentration measurement, is useful.

  The detection of microorganisms was realized quickly and automatically at a low cost.

The usefulness of DOX ^™ was confirmed by inoculating three bacterial species prepared in advance in apheresis (blood component fraction collection) platelet preparations. Serratia marcescens, Staphylococcus aureus (Staphylococcus aureus), Staphylococcus epidermidis (Staphylococcus epidermidis) is inoculated into a platelet preparation derived from a single donor (single blood donor) from Japan Red (10 ⁰ , 10 ¹ , 10 ² CFU / mL) (CFU is an abbreviation for colony forming unit) and cultured at room temperature (20-24 ° C.) for 48 hours. Platelet preparations need to be stored at room temperature in order to retain their platelet functions. Immediately after bacterial inoculation, 1 mL of a sample was taken from the bag and dispensed into a cell of a dissolved oxygen measuring device (DOX-30F), and the dissolved oxygen concentration in the preparation was measured continuously at 35 ° C. for 24 hours. The platelet preparation content of the same lot without bacterial inoculation was set in the apparatus in the same manner as 1 mL, and used as a negative control. The determination of dissolved oxygen consumption was made when a change in the slope was observed. Fig. 1 is a graph showing the results of continuous measurement of dissolved oxygen concentration for 24 hours immediately after inoculation of bacteria into a blood transfusion preparation. Table 1 shows the results of counting the number of bacteria immediately after inoculation of blood transfusion preparations and after incubation at 20-24 ° C for 48 hours. Indicates.

When the test platelet preparation contaminated with S. marcescens (10 ² CFU / mL) was used for measuring dissolved oxygen concentration, a slope indicating dissolved oxygen consumption appeared on the graph and was detectable at 760 minutes. At the same time, 1 mL of the test platelet preparation was collected after incubation at room temperature for 48 hours and the number of bacteria was measured by a conventional method, and 10 ⁷ CFU / mL was confirmed. 10 ¹ CFU / mL was not detectable within 24 hours. The number of bacteria was not detected by the conventional method after 48 hours. Probably the inoculum was killed by the bactericidal ability of the platelet preparation. Test platelet preparations contaminated with S. aureus (10 ¹ , 10 ² CFU / mL) were detectable at 1200 minutes and 500 minutes, respectively, using the dissolved oxygen concentration measurement method (Graph 1). In all the conventional methods after 48 hours, the number of bacteria was 10 ⁷ CFU / mL. In S. epidermidis, neither the oxygen electrode method nor the bacterial count method was detectable at any inoculation concentration of 10 ¹ , 10 ² CFU / mL.

If S. marcrscens, S. aureus was present as 10 ² CFU / mL contamination during the production of the platelet preparation, it is considered that the presence of bacterial contamination can be detected within about 20 hours. The contaminated preparation was stored at room temperature, and the number of bacteria reached 10 ⁷ CFU / mL after 48 hours, which is a general standard time for blood transfusion. When recipients are transfused with this contaminated product, they will experience severe bacteremia. In S. epidermidis, it was not detected by both dissolved oxygen concentration measurement method and bacterial count measurement method, and the inoculum was thought to have disappeared due to the bactericidal ability of fresh blood. From the above, using the dissolved oxygen concentration measurement method, it is possible to detect about 20 hours later by sampling a portion 24 hours after manufacture, which is the time of platelet production or delivery to the drug product use facility, and continuously measuring the dissolved oxygen concentration. It was considered. As a result, there is a high possibility that an accident during blood transfusion can be avoided. This device has already been commercialized as a device that can quickly detect bacterial contamination of food, and is useful as a device for detecting bacterial contamination of blood products, including autotransfusion as a rapid, automated, low-cost practical system. It was considered.

  As an indicator for detecting microbial contamination, there is a high possibility of avoiding accidents during blood transfusion. Detection of microbial contamination of blood products is useful, including autologous blood transfusion using dissolved oxygen concentration measurement method, which is a rapid, automated and low-cost practical system.

Graph showing the results of continuous measurement of dissolved oxygen concentration for 24 hours immediately after inoculation of the blood transfusion product

Claims (2)

  A method for measuring the amount of dissolved oxygen in a transfusion product and measuring the microbial contamination of the transfusion product using the amount as an indicator   A method to detect the slope of the dissolved oxygen concentration consumption curve in transfusion products as an indicator of microbial contamination of transfusion products

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