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目的 :建立简易、准确的诊断内脏利什曼病和病原体鉴定技术。方法 :采用作者设计的杜氏利什曼原虫 ( L.d.)种特异引物 和 ,经 PCR扩增样品内病原体 k DNA2 97bp片段 ,检测 2 2例确诊内脏利什曼病 ( VL )患者骨髓、血、血清共 55份样品和 4例临床疑诊为黑热病患者的骨髓 (共 2 6例 ,59份样品 )。结果 :( 1) PCR法与骨髓涂片镜检符合率为 96.2 % ( 2 5/2 6) ;( 2 ) PCR法检测骨髓、血及血清的总阳性率为 95.4 % ( 2 1/2 2 ) ;( 3) PCR法检测 2 2例骨髓、 16例血样品及 17例血清样品 ,阳性率分别为 91% ( 2 0 /2 2 )、 68.8% ( 11/16)及 2 9.4 % ( 5/17)。 15例骨髓对照样品得自 9例白血病患者及 6例健康人。血及血清对照样品各得自 5例健康人。全部对照未见扩增产物 ,均为阴性。结论 :采用引物 和 进行 PCR扩增检测血内 k DNA特异片段诊断内脏利什曼病有较好前景。
Objective: To establish a simple and accurate diagnosis of visceral leishmaniasis and identification of pathogens. Methods: The specific primers of Leishmania donovani (Ld) were designed and used to amplify the kappa DNA2 97bp fragment of the pathogen in the samples. The bone marrow, serum and serum of 22 patients with visceral leishmaniasis (VL) were detected by PCR. A total of 55 samples and 4 cases of clinical suspected melanoma patients with bone marrow (a total of 26 cases, 59 samples). Results: (1) The coincidence rate of PCR and bone marrow smears was 96.2% (2 5/2 6). (2) The total positive rate of bone marrow, blood and serum detected by PCR was 95.4% (2 1/2 2 ). (2) The bone marrow, 16 blood samples and 17 serum samples were detected by PCR. The positive rates were 91% (20/22), 68.8% (11/16) and 2 9.4% / 17). Fifteen bone marrow control samples were obtained from 9 leukemia patients and 6 healthy individuals. Blood and serum control samples were obtained from 5 healthy individuals. No amplification products were found in all controls, all of which were negative. Conclusion: It is a good prospect to diagnose visceral leishmaniasis by using kp DNA primers and PCR amplification.