Meredith S E, Zijlstra E E, Schoone G J, Kroon C C, van Eys G J, Schaeffer K U, el-Hassan A M, Lawyer P G
Royal Tropical Institute, Amsterdam, The Netherlands.
Arch Inst Pasteur Tunis. 1993 Jul-Oct;70(3-4):419-31.
Detection, diagnosis and identification of Leishmaniasis may be difficult owing to low numbers of parasites present in clinical samples. The PCR has improved the sensitivity and specificity of diagnosis of several infectious diseases. A leishmania specific PCR assay was developed based on the SSUrRNA genes which amplifies DNA of all Leishmania species. Point mutations occurring within the rRNA genes allow differentiation of the Leishmania complexes using primers constructed with the 3/ ends complementary to the specific point mutations present in the SSU rRNA genes of the Leishmania species. Biopsy material, blood, lesion impressions and blood spots on filter paper can be used in the assay. In a longitudinal study on the incidence rates of VL, subclinical cases and PKDL in an endemic region of Sudan, filter paper blood spots from proven and suspected VL patients, PKDL and control samples from an endemic region in Sudan are being taken. The blood spots were analyzed in the DAT and by PCR and results compared with clinical and parasitological data. The first results indicate that the PCR on blood spots is a simple and sensitive means of detecting active VL; in PKDL patients parasites are detectable in the skin.
由于临床样本中存在的寄生虫数量较少，利什曼病的检测、诊断和鉴定可能会很困难。聚合酶链反应（PCR）提高了几种传染病诊断的敏感性和特异性。基于小亚基核糖体RNA（SSUrRNA）基因开发了一种利什曼原虫特异性PCR检测方法，该方法可扩增所有利什曼原虫物种的DNA。rRNA基因内发生的点突变使得利用与利什曼原虫物种SSU rRNA基因中存在的特定位点突变互补的3′端构建的引物能够区分利什曼原虫复合体。活检材料、血液、病变印记和滤纸上的血斑均可用于该检测。在苏丹一个流行地区对内脏利什曼病、亚临床病例和皮肤利什曼病后皮肤利什曼病发病率的纵向研究中，正在采集来自苏丹流行地区确诊和疑似内脏利什曼病患者、皮肤利什曼病后皮肤利什曼病患者以及对照样本的滤纸血斑。对血斑进行直接凝集试验（DAT）和PCR分析，并将结果与临床和寄生虫学数据进行比较。初步结果表明，对血斑进行PCR检测是检测活动性内脏利什曼病的一种简单且敏感的方法；在皮肤利什曼病后皮肤利什曼病患者的皮肤中可检测到寄生虫。
