LAL Test에서 위양성을 나타내는 원인들에 대한 고찰

Consideration About the Bacterial Endotoxin Test Showing False Positive Test Result When Performing LAL Test

환자 몸에 투여되는 주사용제재으로서 약품 내 무균성이 요구되어진다. 현재 대부분의 방사성약품합성 실에서는 사용하기가 비교적 간편하고 민감도가 우수한 Bacterial Endotoxin Test로 방사성약품 내 발열성물질이 존재하는지의 유무를 실험한다. 그렇지만 테스트결과를 해석할 때 가끔씩 위 양성의 결과를 나타내서 결과를 해석 시 혼란을 줄 때도 있다. 이 위 양성의 원인을 알아보기 위해 pH영역이 다른 buffer와 Ethanol sample을 사용해서 실험을 해보았다. 그 결과 pH영역이 너무 산성이나 염기성인 1.0~5.0 그리고 11.2~12.6에서 위 양성의 결과를 나타냈고 Ethanol sample에 있어서는 20~50%에서는 위 양성의 결과를 100%에서는 완전한 양성의 결과를 나타내었다. Pyrogen은 면역력이 약한 환자들에게 있어서 두통이나 발열, 백혈구감소증 등의 증상을 일으키고 심지어 합병증으로 인해 환자들에게 치명적인 반응을 일으키기도 한다. 그러므로 우리는 위양성의 결과에 영향 미치는 이런 요인들을 최대한 배제함으로써 객관적이고 신뢰할만한 Pyrogen test결과를 도출하는게 필요하다.

Purpose: Since radiopharmaceuticals are intended for human administration, it is imperative that we should undergo quality control very strictly. Now almost all the PET laboratories have adopted Bacterial Endotoxin Test as the stand quality control method to monitor whether pyrogen is free or not in the product vial containing crude solution. The aim of this study is to find out the reason why false positive result is observed when using commercially available test vial. Materials and Methods: For this experiment, we used commercially available single test kit (Associates of Cape Code. Inc. USA) and we made pH samples by mixing each buffer whose pH ranges are 1.0 to 12.0. Otherwise we made Ethanol samples diluted with distilled water. After making test samples, it added 0.2 mL to the test vial. Assay mixture in the test vial was incubated in a water bath (Chang Shin Co. KOR) for 60 min at $35{\pm}2^{\circ}C$. Results: After incubation period ($60{\pm}1^{\circ}C$), we inverted the test vial about $180^{\circ}$ To know what pH and how many percentage of Ethanol (Fisher Scientific Korea. Ltd) will affect the reaction. With pH buffer, false positive result was observed at pH 1.0 to 5.0 and 7.7 to 12.6 but at pH 5.2 to.7.5, the test results show negative. It's very strange that we couldn't observe negative test result with Tris buffer at pH 8.4, 8.6, 8.8, 9.0. in other case Ethanol, the test result was seen with 5 to 10% Ethanol. But to my surprise we could see very thick gel formation with 100% Ethanol. Conclusions: In this study, we could notice that pH which is too much acidic or alkalic or high concentrated Ethanol would affect Bacterial Endotoxin Test result. As you know, LAL test is sensitive and very reliable method. Therefore, we are needed to elicit the accurate test result as possible as we can.