TITLE:
Monoclonal antibody technology applied to the detection of Cryptosporidium parvum oocysts in human and cattle fecal samples
AUTHORS:
Vera Codices, Olga Matos, Carlos Novo
KEYWORDS:
Hybridomas; Cryptosporidium parvum; Immunofluorescence; Detection
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.4 No.4A,
April
29,
2013
ABSTRACT: With the discovery that the coccidian parasite Cryptosporidium sp. can cause
severe symptoms in humans, many diagnostic
techniques were quickly implemented such
as microscopic visualization, immunofluorescence and PCR. Currently,
there is no effective drug treatment and none of the current diagnostic
methods is 100% accurate. In this study, a BALB/C mouse was subcutaneously immunized with Cryptosporidium parvum oocysts extract. The spleen was removed and the splenocytes were fused with
SP2/0 myeloma cells in order to obtain hybridoma cells secreting antibodies
specific to C. parvum antigens. Human and cattle fecal samples previously
characterized by microscopy [Ziehl-Neelsen staining (ZN) and Lugol] and PCR for
the presence of C. parvum and Giardia duodenalis, were
analyzed by indirect immunofluorescence, using the developed hybridomas
supernatants. The study shows that the selected hybridomas supernatants
identify C. parvum oocysts in fecal samples in correlation with C. parvum oocysts identified using ZN/PCR. No false positive results were obtained and
the two best supernatants gave 20% -30% of false negative results. No cross
reaction with G. duodenalis was observed. By comparing our results with those obtained
with an immunofluorescence commercial kit, it suggests the potential use of
the monoclonal antibodies present in two of the hybridomas supernatants as a
detection tool of C. parvum. With a reliability of 80.8% and
73.1% versus ZN and PCR methods for IFI, compared with a reliability of 76.9%
and 92.3% versus ZN and PCR for commercial DIF kit, the supernatant
4.1D5 seems to be the most promising subject to further study its usefulness
for C. parvum detection.