TY - JOUR
T1 - Flow cytometric detection of Cryptosporidium oocysts in human stool samples
AU - Valdez, Luis M.
AU - Dang, Han
AU - Okhuysen, Pablo C.
AU - Chappell, Cynthia L.
PY - 1997/8
Y1 - 1997/8
N2 - Cryptosporidium parvum is an important pathogen that causes diarrhea in virtually all human populations. Improved diagnostic methods are needed to understand the risk factors, modes of transmission, and impact of cryptosporidiosis. In the present study, we fluorescently labeled and counted C. parvum oocysts by flow cytometry (FC) and developed a simple and efficient method of processing human stool samples for FC analysis. Formed stool (suspended in phosphate-buffered saline) from an asymptomatic, healthy individual was seeded with known concentrations of oocysts, and oocysts were labeled with a cell wall-specific monoclonal antibody and detected by FC. The method described herein resulted in a mean oocyst recovery rate of 45% ± 16% (median 42%), which consistently yielded a fourfold increase in sensitivity compared to direct fluorescent-antibody assay of seeded stool samples. However, in many instances, FC detected as few as 103 oocysts per ml. Thus, FC provides a reproducible and sensitive method for C. parvum oocyst detection.
AB - Cryptosporidium parvum is an important pathogen that causes diarrhea in virtually all human populations. Improved diagnostic methods are needed to understand the risk factors, modes of transmission, and impact of cryptosporidiosis. In the present study, we fluorescently labeled and counted C. parvum oocysts by flow cytometry (FC) and developed a simple and efficient method of processing human stool samples for FC analysis. Formed stool (suspended in phosphate-buffered saline) from an asymptomatic, healthy individual was seeded with known concentrations of oocysts, and oocysts were labeled with a cell wall-specific monoclonal antibody and detected by FC. The method described herein resulted in a mean oocyst recovery rate of 45% ± 16% (median 42%), which consistently yielded a fourfold increase in sensitivity compared to direct fluorescent-antibody assay of seeded stool samples. However, in many instances, FC detected as few as 103 oocysts per ml. Thus, FC provides a reproducible and sensitive method for C. parvum oocyst detection.
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U2 - 10.1128/jcm.35.8.2013-2017.1997
DO - 10.1128/jcm.35.8.2013-2017.1997
M3 - Article
C2 - 9230372
AN - SCOPUS:0030809236
SN - 0095-1137
VL - 35
SP - 2013
EP - 2017
JO - Journal of Clinical Microbiology
JF - Journal of Clinical Microbiology
IS - 8
ER -