摘要Objective: To explore the feasibility of enzyme-linked immunosorbent assay (ELISA) in detecting syphilis dry blood spots. Methods: Based on dry blood spot samples, laboratory linear dial, laboratory basic dial, laboratory interference dial, and laboratory precision dial were constructed. The linear range, sensitivity, specificity, precision and other performances of ELISA for detecting syphilis dry blood spot samples were comprehensively evaluated, and the stability of dry blood spot samples at 37℃ was detected. In addition, 112 suspected syphilis antibody-positive plasma samples were selected as the control, and dry blood spot samples were prepared accordingly. The clinical sensitivity, clinical specificity, positive predictive value and negative predictive value of dry blood spot samples and plasma samples in ELISA for syphilis detection were compared, and the consistency and correlation between the two samples were analyzed by the Kappa consistency test and Spearman rank correlation analysis. Results: The results of the linear analysis showed that the serial dilution of dry blood spot samples in ELISA for syphilis antibody ranged from 23 to 27, and there was a good linear range [R2=0.9862(P<0.05)]. Sensitivity and specificity analysis showed that the positive coincidence rate and negative coincidence rate of the two detection methods were 100% (15/15). The results of the interference dial test showed that ELISA based on dry blood spot samples could accurately detect 6 syphilis antibody samples from 18 samples, and the detection accuracy rate was 100.00% (6/6). The results of the precision test showed that the RSD of syphilis antibody detection in dry blood spot samples with different dilution times (23, 25 and 27) was 0.24% to 3.87% between spots, 0.06% to 4.07% between batches and 0.49% to 3.88% between days. Within 7 days, the inter-day RSD of dry blood spots with different dilution times (23,25,27) were 0.27%, 0.65% and 0.95%, respectively. The clinical sensitivity, clinical specificity, positive predictive value and negative predictive value of dry blood spot samples in ELISA detection of syphilis antibody were 96.00%(72/75), 100.00%(37/37), 100.00%(72/72) and 92.50%, respectively. The results of the Kappa consistency test and Spearman rank correlation analysis showed that the Kappa value of the two methods was 0.941(P<0.01), and the correlation coefficient ρ between the S/CO ratios of the two methods was 0.211 (P<0.01). The comparison of S/CO ratio results showed that the distribution characteristics of the S/CO ratio between the two methods were similar, and the ratio distribution was relatively concentrated. Conclusion: using dry blood spot samples to perform ELISA for syphilis detection has good precision, strong anti-interference ability and excellent stability. Although false-positive results appear in weak positive samples, it still has a high application value in ELISA for syphilis antibody detection, which can provide an important reference for disease diagnosis.
Abstract：Objective: To explore the feasibility of enzyme-linked immunosorbent assay (ELISA) in detecting syphilis dry blood spots. Methods: Based on dry blood spot samples, laboratory linear dial, laboratory basic dial, laboratory interference dial, and laboratory precision dial were constructed. The linear range, sensitivity, specificity, precision and other performances of ELISA for detecting syphilis dry blood spot samples were comprehensively evaluated, and the stability of dry blood spot samples at 37℃ was detected. In addition, 112 suspected syphilis antibody-positive plasma samples were selected as the control, and dry blood spot samples were prepared accordingly. The clinical sensitivity, clinical specificity, positive predictive value and negative predictive value of dry blood spot samples and plasma samples in ELISA for syphilis detection were compared, and the consistency and correlation between the two samples were analyzed by the Kappa consistency test and Spearman rank correlation analysis. Results: The results of the linear analysis showed that the serial dilution of dry blood spot samples in ELISA for syphilis antibody ranged from 23 to 27, and there was a good linear range [R2=0.9862(P<0.05)]. Sensitivity and specificity analysis showed that the positive coincidence rate and negative coincidence rate of the two detection methods were 100% (15/15). The results of the interference dial test showed that ELISA based on dry blood spot samples could accurately detect 6 syphilis antibody samples from 18 samples, and the detection accuracy rate was 100.00% (6/6). The results of the precision test showed that the RSD of syphilis antibody detection in dry blood spot samples with different dilution times (23, 25 and 27) was 0.24% to 3.87% between spots, 0.06% to 4.07% between batches and 0.49% to 3.88% between days. Within 7 days, the inter-day RSD of dry blood spots with different dilution times (23,25,27) were 0.27%, 0.65% and 0.95%, respectively. The clinical sensitivity, clinical specificity, positive predictive value and negative predictive value of dry blood spot samples in ELISA detection of syphilis antibody were 96.00%(72/75), 100.00%(37/37), 100.00%(72/72) and 92.50%, respectively. The results of the Kappa consistency test and Spearman rank correlation analysis showed that the Kappa value of the two methods was 0.941(P<0.01), and the correlation coefficient ρ between the S/CO ratios of the two methods was 0.211 (P<0.01). The comparison of S/CO ratio results showed that the distribution characteristics of the S/CO ratio between the two methods were similar, and the ratio distribution was relatively concentrated. Conclusion: using dry blood spot samples to perform ELISA for syphilis detection has good precision, strong anti-interference ability and excellent stability. Although false-positive results appear in weak positive samples, it still has a high application value in ELISA for syphilis antibody detection, which can provide an important reference for disease diagnosis.
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