Kaushal Deep C, Kaushal Nuzhat A
Central Drug Research Institute, Lucknow, India.
Immunol Invest. 2002 May;31(2):93-106. doi: 10.1081/imm-120004801.
We have previously demonstrated, using polyclonal and monoclonal antibodies, that the lactate dehydrogenase (LDH) of malaria parasites is immunologically distinct from the host enzyme. The polyclonal antibodies, produced against the affinity purified plasmodial LDH (pLDH) in rabbits, showed specificity to LDH of malaria parasites. In the present study, these anti-pLDH polyclonal antibodies were used to develop an immunodiagnostic test (immunodot enzyme assay of plasmodial LDH) based on the detection of parasite LDH in patient blood. The immunodot enzyme assay of plasmodial LDH was evaluated using blood samples from patients with malaria or other infections. Out of 502 microscopically positive malaria blood samples, 497 blood samples showed positive immunodot assays of pLDH while all the 423 microscopically negative cases were found negative by our test. The blood samples from other infections and non-endemic controls were negative by the immunodot enzyme assay of pLDH. This LDH based test was also found negative in blood samples of cured patients 7 days after chloroquine treatment. The test is simple to perform, can be read visually, econimal, highly specific with a sensitivity of approximately 99% and is thus suitable for accurate diagnosis of malaria in field conditions.
我们之前利用多克隆抗体和单克隆抗体证明,疟原虫的乳酸脱氢酶(LDH)在免疫方面与宿主酶不同。针对兔体内亲和纯化的疟原虫LDH(pLDH)产生的多克隆抗体,对疟原虫的LDH具有特异性。在本研究中,这些抗pLDH多克隆抗体被用于开发一种免疫诊断测试(疟原虫LDH免疫斑点酶测定法),该方法基于检测患者血液中的寄生虫LDH。使用疟疾患者或其他感染患者的血样对疟原虫LDH免疫斑点酶测定法进行评估。在502份经显微镜检查呈阳性的疟疾血样中,497份血样的pLDH免疫斑点测定呈阳性,而所有423份经显微镜检查呈阴性的病例经我们的检测均为阴性。来自其他感染患者和非流行地区对照的血样经pLDH免疫斑点酶测定呈阴性。在氯喹治疗7天后的治愈患者血样中,该基于LDH的检测也呈阴性。该检测操作简单,可目视读取结果,经济实惠,特异性高,灵敏度约为99%,因此适用于在现场条件下准确诊断疟疾。