中国寄生虫学与寄生虫病杂志 ›› 2018, Vol. 36 ›› Issue (4): 366-369.

• 论著 • 上一篇    下一篇

一例输入性非洲锥虫病的实验室诊断

林耀莹, 张山鹰*(), 谢汉国, 欧阳榕, 陈朱云, 刘庆生   

  1. 福建省疾病预防控制中心,福州 350001
  • 收稿日期:2018-01-18 出版日期:2018-08-30 发布日期:2018-09-06
  • 通讯作者: 张山鹰
  • 基金资助:
    福建省科技计划引导性项目(No. 2016Y0009)

Laboratory diagnosis of an imported case of African trypanosomiasis

Yao-ying LIN, Shan-ying ZHANG*(), Han-guo XIE, Rong OUYANG, Zhu-yun CHEN, Qing-sheng LIU   

  1. Fujian Provincial Center for Disease Control and Prevention, Fuzhou 350001, China
  • Received:2018-01-18 Online:2018-08-30 Published:2018-09-06
  • Contact: Shan-ying ZHANG
  • Supported by:
    Supported by Fujian Science and Technology Plan Guided Project (No. 2016Y0009)

摘要:

目的 对一例输入性非洲锥虫病进行实验室诊断。方法 收集住院患者病例资料,采用吉氏染色法镜检患者的血样和脑脊液样品。提取患者血样中布氏锥虫基因组DNA,以布氏锥虫表达位点相关基因(ESAG)、冈比亚锥虫特异性糖蛋白(TgsGP)和罗得西锥虫血清抗性相关(SRA)基因为靶基因,进行PCR扩增,对扩增产物进行电泳、测序,在GenBank中进行同源性比对。结果 患者,女,41岁,福建福州人, 在坦桑尼亚赛伦盖蒂国家公园观看动物期间(2018/07/28-29),右后脚跟被不明昆虫叮咬,8月6日回国。8月8日开始出现发热、咳嗽、全身无力等症状,于福州市某大型三甲医院就诊,临床诊断为感染性发热。经头孢地尼(抗感染)和散利痛(退热)治疗后,未见好转。患者于8月9日转诊到福州市另一大型三甲医院,经头孢唑肟(抗感染),达菲(抗病毒),沐舒坦、十位龙胆花(化痰),天兴、复方二氯醋酸异丙胺(保肝),营养支持等治疗,体温仍控制不佳,肝酶进行性升高。8月14日,患者将医院采集的血样和脑脊液送福建省疾病预防控制中心检测疟原虫。吉氏染色镜检结果显示,全血样本中未见疟原虫,但可见疑似布氏锥虫的锥鞭毛体。脑脊液中未检出锥虫。对患者血样作进一步分子生物学检测,PCR结果显示,布氏锥虫ESAG属特异性引物扩增片段长度为260 bp,冈比亚锥虫特异性引物未扩增出条带,罗得西锥虫特异性SRA引物扩增片段长度为266 bp。序列比对结果显示,EASG的扩增序列与AY682003.1同源性为95%,SRA基因的扩增序列与AJ345058.1同源性为99%。患者确诊为罗得西锥虫感染后,分别于第1、3、7、14、21天静脉注射20 mg/kg苏拉明(由WHO提供),每次注射剂量不超过1 g。1个疗程结束后,患者基本恢复,出院后按医嘱定期至医院复查,均未检出锥虫。结论 该病例经实验室诊断分析,确诊为输入性罗得西锥虫感染。

关键词: 罗得西锥虫, 非洲锥虫病, 输入性, 诊断

Abstract:

Objective To diagnose an imported case of suspected African trypanosomiasis by laboratory tools. Methods Clinical information of the patient was collected. The blood and cerebrospinal fluid samples were examined by microscopy after Giemsa staining. DNA of Trypanosoma brucei was extracted from the patient blood sample, and amplified by PCR using primers for T. brucei genus expression site-associated gene(ESAG), T. brucei gambiense-specific glycoprotein(TgsGP) and T. brucei rhodesiense serum resistance-associated (SRA) gene. The PCR products underwent electrophoresis, sequencing, and the sequences were blasted in GenBank. Results The patient was a 41-year-old female from Fuzhou, Fujian Province. She was bitten by an unknown insect in Serengeti National Park of Tanzania (July 28-29, 2018). After returning to China on August 6, she began to develop symptoms of fever, fatigue, and coughing on August 8. She visited a class A hospital in Fuzhou City, where he was disgnosed with infectious fever. The symptoms were not improved after treatment with efdinir (for anti-infection) and Saridon (for fever). The patient was transferred to another class A hospital in the city on August 9, but the fever was still not effectively ameliorated after a combination of anti-infective eftizoxime, anti-viral Tamiflu, phlegm-resolving Mucosolvin and Shiwei Longdanhua Capsule, liver-protective Tianxing and diisopropylamine chloroacetate, and nutrition support, accompanied by progressive increases of liver enzymes. On August 14, the blood and cerebrospinal fluid samples of the patient were sent to Fujian Provincial Center for Disease Control and Prevention for Plasmodium examination. Microscopy of the Giemsa-stained samples showed suspected African trypanosome trypomastigotes in the blood sample, but failed to find Plasmodium. No Trypanosoma was found in the cerebrospinal fluid. PCR results showed a product of 260 bp with the primers for ESAG, no specific product with the primers for TgsGP but a product of 266 bp with the primers for SRA. Results of sequence alignment revealed that the EASG amplified sequence had a 95% homology with AY682003.1, and the SRA amplified sequence had a 99% homology with AJ345058.1. After confirmation of T. brucei rhodesiense infection, the patient was intravenously injected with 20 mg/kg suramin (provided by WHO) on days 1, 3, 7, 14 and 21, with dosage no more than 1 g at each injection. After one treatment course, the patient basically recovered, and was not detected with Trypanosoma at regular checkups in the hospital. Conclusion The case is confirmed to be T. brucei rhodesiense infection by the laboratory diagnosis.

Key words: Rhodesiense, African trypanosomiasis, Imported, Diagnosis

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