AlTameemi, Abeer Muataz Abed
(2018)
Comparison of the efficiencies of customized CSA-and rlectin-elisas for detection of anti-amoebic antibody based on selected aborigines serum samples.
Masters thesis, Universiti Sains Malaysia.
Abstract
Orang Asli, the Malaysian aborigines live in remote jungles, are endemic for amoebiasis.
Understanding the disease prevalence and distribution is important for the control and
preventive measures. A major indicator that reflects on the disease incidence is the
presence of highly elevated anti-amoebic antibody. However, continuous screening of
the antibody using commercial kits is impractical due to the high cost. Cheaper but valid
in-house customized assay should be employed instead to attain cost-effective and
sustainable outcome. Hence, the presence study aimed to compare the efficiencies of
customized crude soluble antigen (CSA)-ELISA and recombinant lectin (rLectin)-
ELISA for detection of anti-amoebic antibody in selected Orang Asli serum samples,
which were prior defined by commercial indirect haemagglutination assay (IHA). CSA
was produced from axenically grown Entamoeba histolytica trophozoites, while rLectin
was overexpressed and purified from Escherichia coli harbouring the corresponding
recombinant gene. For the development of the two customized ELISAs, antigen
concentration, serum and conjugated antibody dilutions were optimized. The
efficiencies of both customised ELISAs were determined by 30 positive and negative
serum samples pre-determined by commercial IHA. According to receiver operating
characteristics (ROC) curve analysis, both customized ELISAs showed excellent
diagnostic performances. As compared to the reference assay, the positive percentage
agreement of CSA-ELISA (91%) was higher than that of rLectin-ELISA (61%), whileboth ELISAs showed the same negative percentage agreement, which was 97%.
According to analysis of ELISA ODs, the positive delta value for CSA-ELISA (0.4472)
was higher than that of rLectin-ELISA (0.2673), while the negative delta value of
rLectin-ELISA (-1.209) was lower than that of CSA-ELISA (-1.175). In conclusion, this
study demonstrated that CSA-ELISA was more sensitive in detecting the presence of
anti-amoebic antibody, while rLectin-ELISA was more specific in ruling out negative
cases.
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