Makler M T, Hinrichs D J
Clinical Pathology and Research Service, Veterans Administration Medical Center, Portland, Oregon.
Am J Trop Med Hyg. 1993 Feb;48(2):205-10. doi: 10.4269/ajtmh.1993.48.205.
This report describes an enzyme assay for the detection of Plasmodium falciparum. The assay is based on the observation that the lactate dehydrogenase (LDH) enzyme of P. falciparum has the ability to rapidly use 3-acetyl pyridine NAD (APAD) as a coenzyme in the reaction leading to the formation of pyruvate from lactate. Human red blood cell LDH carries out this reaction at a very slow rate in the presence of APAD. We measured the development of APADH and found that the formation of this product could establish the basis of an assay that detected the presence of P. falciparum from in vitro cultures at parasitemia levels of 0.02%. We also had occasion to use this assay with clinical samples. We found a correlation between levels of parasitemia and the activity of parasite LDH. Parasite LDH (pLDH) activity could be measured in blood hemolysates and in plasma and serum from patients with malaria. We used the serum assay for pLDH and followed the level of pLDH in a patient with cerebral malaria prior to antimalarial treatment and during the recovery period. From these initial studies, it is evident that the measurement of pLDH has a correlation with parasitemia and may offer a method that can be developed into a simple test for the detection of Plasmodium parasitemia.
本报告描述了一种用于检测恶性疟原虫的酶检测方法。该检测方法基于以下观察结果:恶性疟原虫的乳酸脱氢酶(LDH)能够在由乳酸形成丙酮酸的反应中迅速将3-乙酰吡啶NAD(APAD)用作辅酶。在APAD存在的情况下,人类红细胞LDH进行此反应的速度非常慢。我们测量了APADH的生成,发现该产物的形成可以为一种检测方法奠定基础,该方法能够检测体外培养物中疟原虫血症水平为0.02%时恶性疟原虫的存在。我们也有机会将此检测方法用于临床样本。我们发现疟原虫血症水平与寄生虫LDH的活性之间存在相关性。寄生虫LDH(pLDH)活性可以在疟疾患者的血液溶血产物以及血浆和血清中进行测量。我们使用血清检测pLDH,并在抗疟治疗前和恢复期跟踪一名脑型疟疾患者的pLDH水平。从这些初步研究中可以明显看出,pLDH的测量与疟原虫血症相关,并且可能提供一种可以发展成为检测疟原虫血症的简单测试方法。