Development And Performance Evaluation Of A Dry Chemical Enzymatic Method Multitest Kit For Bacterial Vaginosis

Bacterial vaginal(BV) is a common disease of obstetrics and gynecology, which is much harmful for women. Quick and efficient diagnosis of BV is important before gynecology, obstetrics surgical and abortion. Samples of BV are vaginal secretions, whose composition is complicated. BV is not infected by a single bacterial but a group of anaerobic bacterial, which is difficult for BV test. Currently, there is few of efficient and effective products for BV diagnosis. BV blue that developed abroad, used enzymatic method detecting BV and solve the problem. But its high cost, false negative, complex operation and so on factors affected the popularization and application in our country. In order to meet the domestic demand for rapid detection of BV, we studied one bacterial vaginal disease kits by dry chemical enzymatic, which had good sensitivity, high degree of specific, convenient operating and low costs, which would apply to the BV diagnosis. Methods and results:1. The main material: designed one special reaction plate, color card. Filtered carrier, substrates and quality control, and formulated the relevant quality standards.2. The main technological research: selected 4-Aminoantipyrine, sialic acid substrates B, L-proline paranitroaniline, bromocresol green as hydrogen peroxide, saliva acid glycosides enzyme, proline aminopeptidase and pH indicators substrates, Through a series of orthogonal experiment and optimization for each index of the optimal formula, flow diagram of the product is formed.3. The reaction system research: the samples of this kit are vaginal secretions, and the best sample amount is 30μl, the reaction at 37 for 15 minutes, and read the ℃result after incubation for 5 minutes.4. Analysis of the performance study: analysised the limit, specificity, accuracy, repeatability, and assessment of three batches kits, and meet the requirements.5. Stability studies: the kit performance was stable and could meet the requirements accelerated at 37 ℃for 9 days. The performance could meet the requirements to simulate the transport test at 37 ℃for 7 days. Its performance was stable in 12 months under conventional storage conditions. The reaction plate once opened can be stable for four hours. Vaginal secretions samples after collected should be used within an hour.6. Positive standard studies: tested 100 normal samples and 50 positive samples each of hydrogen peroxide, proline amino peptide enzyme and saliva acid glycosides enzyme, using ROC curve method to establish the three indicators of positive judgment standard.7. Clinical research: there is significant difference between BVblue kit and Amsel golden standard in 681 test cases of clinical samples, and the sensitivity of BVblue kit is 85.58%. There is no significant difference between BVblue kit and the index factor sialidase of our kit. The results of our kit using four indicators joint detection compared to Amsel gold standard, the sensitivity was 98.08%, specificity was 99.48%, positive predictive value was 97.14%, negative predictive value was 99.65%, and the total coincidence rate was 99.27%. This kit has better detection performance than BVblue kit. Conclusion: The bacterial vaginal disease joint detection kits self designed and developed can better applied to clinical diagnosis of BV with convenient and quick operation, stable performance, high sensitivity, and good specificity.