Schönian Gabriele, Nasereddin Abedelmajeed, Dinse Nicole, Schweynoch Carola, Schallig Henk D F H, Presber Wolfgang, Jaffe Charles L
Institute of Microbiology and Hygiene, Humboldt University, Charité, Berlin, Germany.
Diagn Microbiol Infect Dis. 2003 Sep;47(1):349-58. doi: 10.1016/s0732-8893(03)00093-2.
Leishmaniasis diagnosis in regions where multiple species exist should identify each species directly in the clinical sample without parasite culturing. The sensitivity of two PCR approaches which amplify part of the ssu rRNA gene and the ribosomal internal transcribed spacer (ITS), respectively, was determined using human and dog blood seeded with Leishmania promastigotes. ssu-rDNA-PCR was more sensitive than ITS1-PCR, however species identification was not possible by the former approach. When a nested ITS1-PCR was used its sensitivity equaled the ssu-rDNA-PCR. Digestion of ITS1 amplicon with the restriction enzyme HaeIII distinguished all medically relevant Leishmania species. ITS1-PCR was used to diagnose 162 local and imported suspected cases of leishmaniasis in Israel, the Palestinian Authority and Germany. 113 cases (69.7%) were positive by PCR and species identification was possible in 110 samples. Leishmania DNA was also amplified and identified at the species level from archived non-stained and Giemsa stained microscope slides.
在存在多种利什曼原虫物种的地区，利什曼病的诊断应在临床样本中直接鉴定出每种物种，而无需进行寄生虫培养。分别使用接种了前鞭毛体利什曼原虫的人血和狗血，测定了两种分别扩增小亚基核糖体RNA（ssu rRNA）基因部分和核糖体内部转录间隔区（ITS）的PCR方法的灵敏度。小亚基核糖体DNA-PCR（ssu-rDNA-PCR）比ITS1-PCR更灵敏，然而，前一种方法无法进行物种鉴定。当使用巢式ITS1-PCR时，其灵敏度与ssu-rDNA-PCR相当。用限制性内切酶HaeIII消化ITS1扩增子可区分所有与医学相关的利什曼原虫物种。ITS1-PCR用于诊断以色列、巴勒斯坦权力机构和德国的162例本地和输入性疑似利什曼病病例。113例（69.7%）PCR检测呈阳性，110份样本能够进行物种鉴定。还从存档的未染色和吉姆萨染色的显微镜载玻片上扩增并在物种水平上鉴定了利什曼原虫DNA。
