Highly specific detection of Cryptosporidium spp. oocysts in human stool samples by undemanding and inexpensive phase contrast microscopy

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• 作者：Ralf Ignatius ; Thomas Klemm ; Steffen Zander ; Jean Bosco Gahutu…
• 关键词：Cryptosporidium ; Phase contrast microscopy ; RT ; PCR ; Rwandan children ; ELISA
• 刊名：Parasitology Research
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：115
• 期：3
• 页码：1229-1234
• 全文大小：394 KB
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• 作者单位：Ralf Ignatius (1) (2)
  Thomas Klemm (1)
  Steffen Zander (3) (5)
  Jean Bosco Gahutu (4)
  Peter Kimmig (1)
  Frank P. Mockenhaupt (3)
  Thomas Regnath (1)
  
  1. Laboratory Enders and Partners, Rosenbergstr. 85, 70193, Stuttgart, Germany
  2. Department of Microbiology and Hygiene, Charité-Universitätsmedizin Berlin, Berlin, Germany
  3. Institute of Tropical Medicine and International Health, Charité-Universitätsmedizin Berlin, Berlin, Germany
  5. Agents of Mycoses, Parasitoses and Mycobacterioses, Robert Koch-Institute, Berlin, Germany
  4. University Teaching Hospital of Butare and School of Medicine and Pharmacy, University of Rwanda, Butare, Rwanda
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Medical Microbiology
  Microbiology
  Immunology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-1955

文摘

To compare phase contrast microscopy (PCM) of unstained slides for the detection of Cryptosporidium spp. oocysts with a commercially available enzyme immunoassay (EIA) for the detection of cryptosporidial antigen in human stool samples, we prospectively analysed by both methods 463 fresh human stool samples obtained from diarrhoeic patients between July and October 2014. Compared with the EIA, the sensitivity, specificity, positive and negative predictive value of PCM were 88.9 % (95 % confidence interval (CI), 66.0–98.1 %), 100 % (95 % CI, 99.0–100 %), 100 % (95 % CI, 77.3–100 %) and 99.6 % (95 % CI, 98.3–100 %), respectively. Additionally, we retrospectively examined with PCM 65 fixed stool samples that had been collected in 2010 from mostly asymptomatic Rwandan children <5 years of age; 14 of these samples had previously yielded positive results with a highly sensitive real-time (RT)-PCR. PCM detected cryptosporidia in 5/14 RT-PCR-positive samples, and notably, also in one of 51 RT-PCR-negative samples, which was subsequently confirmed by acid-fast staining. Positive and negative percent agreement of PCM with RT-PCR were 35.7 % (95 % CI, 16.2–61.4 %) and 98.0 % (95 % CI, 88.7–100 %), respectively. Positive PCM results were associated with higher RT-PCR cycle threshold values (p = 0.044). In conclusion, PCM offers a highly specific, undemanding and inexpensive method for the laboratory diagnosis of acute human cryptosporidiosis independent of the causative Cryptosporidium species.