Riera C, Fisa R, Udina M, Gállego M, Portus M
Laboratori de Parasitologia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona E-08028, Spain.
Trans R Soc Trop Med Hyg. 2004 Feb;98(2):102-10. doi: 10.1016/s0035-9203(03)00015-4.
The extent of cryptic leishmaniasis in blood donors from a Spanish endemic area, (Eivissa Island) was studied using various immunological and parasitological methods. Sera from 656 blood donors were analysed: 16 (2.4%) were positive by ELISA and 50 (7.6%) by Western blot. Peripheral blood mononuclear cells (PBMC) and buffy coat (BC) samples, were analyzed by culture and nested-PCR. DNA of L. infantum was amplified in 27 (22.1%) of 122 PBMC. Parasites were isolated in 3 (4.5%) of 67 BC cultures and the strains were identified as L. infantum zymodeme MON-28. No parasites were isolated in PBMC culture. After 12 months, a second blood sample was obtained from 18 blood donors who were positive by nested-PCR in the first extraction; nine of them remained positive. Delayed type hypersensitivity (DTH) tests on 15/67 donors (22.3%) were positive. Comparison of results obtained by ELISA, WB and DTH; ELISA, WB and nested-PCR and nested-PCR and BC culture showed a significant association (Pearson test, P < 0.05). L. infantum zyodeme MON-28 was identified in three strains isolated from asymptomatic donors, which suggests a low virulence capacity of these strains. The detection of Leishmania DNA in a high number of asymptomatic subjects supports the need to monitor it in blood donors endemic areas.
运用多种免疫学和寄生虫学方法,对西班牙地方病流行区(伊维萨岛)献血者中隐匿性利什曼病的感染程度进行了研究。分析了656名献血者的血清:酶联免疫吸附测定(ELISA)检测出16例(2.4%)呈阳性,蛋白质印迹法检测出50例(7.6%)呈阳性。通过培养和巢式聚合酶链反应(PCR)对外周血单个核细胞(PBMC)和血沉棕黄层(BC)样本进行了分析。在122份PBMC样本中,有27份(22.1%)扩增出婴儿利什曼原虫的DNA。在67份BC培养物中,有3份(4.5%)分离出寄生虫,这些菌株被鉴定为婴儿利什曼原虫酶解模式MON-28。在PBMC培养中未分离出寄生虫。12个月后,从首次提取时巢式PCR呈阳性的1十八名献血者中采集了第二份血样;其中9人仍呈阳性。对15/67名献血者(22.3%)进行的迟发型超敏反应(DTH)试验呈阳性。ELISA、蛋白质印迹法和DTH;ELISA、蛋白质印迹法和巢式PCR;以及巢式PCR和BC培养所获结果的比较显示存在显著相关性(Pearson检验,P<0.05)。从无症状献血者中分离出的三株菌株被鉴定为婴儿利什曼原虫酶解模式MON-28,这表明这些菌株的毒力较低。在大量无症状受试者中检测到利什曼原虫DNA,这支持了在地方病流行区对献血者进行监测的必要性。