C Kittigul

Kasetsart University, Krung Thep, Bangkok, Thailand

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Publications (3)5.23 Total impact

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    ABSTRACT: A modified sandwich enzyme-linked immunosorbent assay using biotin-streptavidin system (BS-ELISA) was developed to determine levels of tumor necrosis factor-alpha (TNF-alpha) in serum samples of children infected with dengue virus (n=99) and healthy controls (n=41). The minimum detectable concentration of TNF-alpha by the BS-ELISA was 3.3 pg/ml. The mean TNF-alpha level was highest in those patients with dengue shock syndrome (DSS) or dengue hemorrhagic fever (DHF) grade III (37.44+/-42.0 pg/ml). Lower levels were found in DHF grade I (28.44+/-42.7 pg/ml), DHF grade II (24. 21+/-25.4 pg/ml) and dengue fever (DF) (14.10+/-24.0 pg/ml). TNF-alpha in the sera of DF and DHF patients could be detected on days 2-6 after the onset of fever, the high level occurring on day 5. TNF-alpha was detected in 41.4% (24.01+/-35.2 pg/ml) of dengue virus infected patients and 7.3% (4.2+/-15.6 pg/ml) of control subjects. The sera of patients contained significantly higher levels of TNF-alpha than the sera of controls, P-value<0.001. DHF patients had significantly higher levels of TNF-alpha than DF patients (P-value=0.020) but no difference in the TNF-alpha levels from sera of DHF grades I-III patients was observed (P-value=0.295). The results indicate that the BS-ELISA is a very sensitive method for determining TNF-alpha in serum samples of DF and DHF patients. The TNF-alpha levels might be associated with dengue virus infection and related to disease severity of DHF.
    Journal of Virological Methods 10/2000; 90(1):51-7. · 1.90 Impact Factor
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    ABSTRACT: A biotin-streptavidin system was adapted to an IgM-capture ELISA for detection of dengue antibodies in human sera. To develop this assay, high titers of antibodies to flavivirus were purified by ion-exchange chromatography (DEAE-cellulose) and labeled with biotin. Heavy chain-specific goat anti-human IgM was first bound to the wells of a polystyrene microtiter plate, followed by binding of IgM in test specimens, and the use of tetravalent dengue antigens (dengue 1-4), biotin-labeled anti-flavivirus IgG, and streptavidin-peroxidase conjugate. The sensitivity and specificity of the IgM-capture biotin-streptavidin ELISA (IgM-BS-ELISA) in acute sera were 83.3% of patients with dengue infection and 95.3% of nondengue-infected cases, respectively. The positive predictive value was 92.4% and the negative predictive value was 89.2%. The efficiency of test was 90.4%. In convalescent sera, the sensitivity and specificity of IgM-BS-ELISA were 100% and 92.6%, respectively. The predictive values of positive and negative results were 90.3% and 100%, respectively. The efficiency of test was 95.6%. The agreement rate of IgM-BS-ELISA and standard hemagglutination inhibition test was good: kappa (kappa) values were 0.79 for acute sera and 0.91 for convalescent sera. The correlation between two methods was quite good, with correlation coefficients (r) of 0.76 for acute sera and 0.85 for convalescent sera (P < 0.001). The results indicate that the IgM-BS-ELISA is highly sensitive, specific, simple to perform, and rapid.
    The American journal of tropical medicine and hygiene 10/1998; 59(3):352-6. · 2.53 Impact Factor
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    ABSTRACT: The presence of dengue virus antigens in acute sera and peripheral blood mononuclear cells (PBMC) from dengue infected patients were determined by a biotin-streptavidin enzyme-linked immunosorbent assay (BS-ELISA). The frequency of the antigens detected in PBMC was higher than that in sera (53.8% vs 18.9%). In comparison with sera, the detection rate in PBMC was greater than six times: 7 cases were positive only in sera whereas 44 cases were positive only in PBMC, p < 0.001. The presence of the antigens in the sera did not depend on the severity of the disease, i.e. dengue fever, dengue hemorrhagic fever (grades I and II) or dengue shock syndrome (grades III and IV). In contrast, the presence of the antigens in PBMC increased from 36.8% to 100% when the infection was more severe. The dengue virus antigens could be detected in the samples collected between day 2 and day 7 after onset of the disease with the highest rate of detection (68.8%) in PBMC collected on day 4. The data suggest the use of PBMC with access to the appropriate acute-phase specimen for detection of dengue virus antigens.
    Asian Pacific journal of allergy and immunology / launched by the Allergy and Immunology Society of Thailand 12/1997; 15(4):187-91. · 0.79 Impact Factor

Publication Stats

39 Citations
5.23 Total Impact Points

Institutions

  • 2000
    • Kasetsart University
      • Department of Microbiology
      Krung Thep, Bangkok, Thailand
    • Mahidol University
      • Department of Microbiology
      Bangkok, Bangkok, Thailand