A flow cytometric protocol for detection of Cryptosporidium

• Type: Article in International Scientific Journal
• Date: 2008

Authors

Joana MM Barbosa (Author) Other; S Costa -de- Oliveira (Author) FMUP; Acacio G Rodrigues (Author) FMUP; Thomas Hancheid (Author) Other; Howard Shapiro (Author) Other; Cidália Pina - Vaz (Author) FMUP

Journal

Title: Cytometry Part A

Vol. 73A

Pages: 44-47

ISSN: 1552-4922

Publisher: Wiley-Blackwell

Indexing

Pubmed / Medline

Scientific classification

FOS: Medical and Health sciences > Other medical sciences

CORDIS: Health sciences

Other information

Abstract (EN): Cryptosporidium parvum is transmitted through water and can cause severe diarrhea. The diagnosis is usually based upon observer-dependent microscopic detection of oocysts, with rather low sensitivity and specificity. Our objective was to optimize a flow cytometric (FC) protocol for the detection of C. parvum. A specific monoclonal antibody conjugated with R-phycoerythrin was incubated with dead oocysts to determine the optimal antibody concentration. Serial concentrations of oocysts were stained with the optimized concentration and analyzed by FC. The lower detection limit was determined, and the possibility of cross-reaction was investigated using prokaryotic and eukaryotic microorganisms. A FC protocol was optimized to detect oocysts in spiked human stools. The optimal antibody concentration was found to be 3.0 mg/ml. The lowest number detectable was 2 × 103 oocysts/ml. Staining procedure was specific, as no cross-reactions were observed. This reliable and easy FC protocol allow the specific detection of Cryptosporidium oocysts, even at very low concentrations, which is important for public health and further studies of treatment efficacy. '

Language: Portuguese

Type (Professor's evaluation): Scientific