Comparison of conventional staining methods and monoclonal antibody-based methods for Cryptosporidium oocyst detection

M. J. Arrowood, Charles R Sterling

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Abstract

The sensitivity and specificity of seven microscopy-based Cryptosporidium oocyst detection methods were compared after application of unconcentrated fecal smears. The seven methods were as follows: (i) a commercial acid-fast (AF) stain (VOLU-SOL) method, (ii) Truant auramine-rhodamine (AR) stain method, (iii) fluorescein-conjugated C1B3 monoclonal antibody (MAb) direct fluorescence method, (iv) OW3 MAb indirect fluorescence method, (v) biotinylated OW3 indirect fluorescence method, (vi) biotinylated OW3-indirect diaminobenzidine (DAB) method, and (vii) biotinylated OW3-aminoethylcarbazole (AEC) method. A total of 281 randomly collected Formalin-fixed fecal samples (submitted to the Maricopa County Health Department, Phoenix, Ariz.) and 30 known positives (Formalin-fixed and K2Cr2O7-preserved stools from our laboratory) were examined in a blind test; 32 of 311 samples (10.3%) were confirmed positive. Of the confirmed positives, 40.6% were identified by the AF method, 93.8% were identified by the AR method, 93.8% were identified by the C1B3 method, 81.3% were identified by the OW3-DAB method, 71.9% were identified by the OW3-AEC method, 100% were identified by the OW3 indirect fluorescence method, and 100% were identified by the biotinylated OW3 indirect fluorescence method. False-positives were encountered by the AF and AR methods (52.0 and 85.7% specificity, respectively), while no false-positives were encountered by the MAb-based methods. Oocysts in infected tissue sections were easily detected by the MAb-based methods.

Original languageEnglish (US)
Pages (from-to)1490-1495
Number of pages6
JournalJournal of Clinical Microbiology
Volume27
Issue number7
StatePublished - 1989

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Cryptosporidium
Oocysts
Monoclonal Antibodies
Staining and Labeling
Benzophenoneidum
Fluorescence
Rhodamines
Formaldehyde
Acids
Coloring Agents
Potassium Dichromate

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

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title = "Comparison of conventional staining methods and monoclonal antibody-based methods for Cryptosporidium oocyst detection",
abstract = "The sensitivity and specificity of seven microscopy-based Cryptosporidium oocyst detection methods were compared after application of unconcentrated fecal smears. The seven methods were as follows: (i) a commercial acid-fast (AF) stain (VOLU-SOL) method, (ii) Truant auramine-rhodamine (AR) stain method, (iii) fluorescein-conjugated C1B3 monoclonal antibody (MAb) direct fluorescence method, (iv) OW3 MAb indirect fluorescence method, (v) biotinylated OW3 indirect fluorescence method, (vi) biotinylated OW3-indirect diaminobenzidine (DAB) method, and (vii) biotinylated OW3-aminoethylcarbazole (AEC) method. A total of 281 randomly collected Formalin-fixed fecal samples (submitted to the Maricopa County Health Department, Phoenix, Ariz.) and 30 known positives (Formalin-fixed and K2Cr2O7-preserved stools from our laboratory) were examined in a blind test; 32 of 311 samples (10.3{\%}) were confirmed positive. Of the confirmed positives, 40.6{\%} were identified by the AF method, 93.8{\%} were identified by the AR method, 93.8{\%} were identified by the C1B3 method, 81.3{\%} were identified by the OW3-DAB method, 71.9{\%} were identified by the OW3-AEC method, 100{\%} were identified by the OW3 indirect fluorescence method, and 100{\%} were identified by the biotinylated OW3 indirect fluorescence method. False-positives were encountered by the AF and AR methods (52.0 and 85.7{\%} specificity, respectively), while no false-positives were encountered by the MAb-based methods. Oocysts in infected tissue sections were easily detected by the MAb-based methods.",
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