ABSTRACT
Objective@#To establish a simple and rapid loop-mediated isothermal amplification (LAMP) to detect Candida albicans (CA) and explore its clinical value.@*Methods@#The Primer Explorer 5.0 software was used to design 4 primers for amplification of CA by LAMP. The system and conditions of LAMP reaction were optimized to evaluate its specificity and the minimum limit in the detection. The vaginal swabs were collected from 123 vulvovaginal candidiasis (VVC) patients and 42 healthy individuals. Fungal culture, LAMP test, PCR test and 1.79 mol/L KOH microscopy were conducted in parallel. Fungal culture was used as the reference method for VVC diagnosis. The positive rates between two groups were compared by Pearson χ<sup>2</sup> test. The consistency of the results from LAMP, PCR, microscopy and culture were analyzed by Kappa test and P＜0.05 was considered as statistically significant difference. ROC curve analysis was used to evaluate LAMP and PCR test for diagnosis of VVC.@*Results@#The optimum reaction temperature of LAMP was 61 ℃ with high specificity. No cross reaction with other strains was found. The minimum detection limit was 10<sup>3</sup> copies/ul. The positive rates of LAMP, PCR and microscopy between VVC and healthy group showed statistically significant difference（LAMP: χ<sup>2</sup>=68.576；PCR: χ<sup>2</sup>=64.918；microscopy: χ<sup>2</sup>=50.076，P＜0.01）. LAMP detection and PCR showed good consistency (κ=0.744, 0.720), but microscopy examination showed poor consistency (κ=0.533). LAMP showed diagnostic sensitivity of 87.62%, specificity of 88.33%, positive predictive value of 92.93% and negative predictive value of 80.30%. The area under the curves of LAMP and PCR were 0.873 and 0.888, respectively. No difference in efficacies between LAMP and PCR was found (Z=0.849, P=0.395 6), but the lowest detection time of LAMP was shorter than 1 hour.@*Conclusion@#A rapid, reliable, sensitive and specific LAMP technique for detecting CA was established. The comprehensive screening performance should be superior to the routine method in laboratories, so LAMP could be used for the supplementary diagnosis and monitoring therapeutic effects in CA infection.
ABSTRACT
@#Objective To observe the effect of Sinisan on the learning and memorial ability and the expression of synaptohydin in the hippocampus of exercise-fatigue rats. Methods Rats were made into the fatigue model by Exhaustive Swimming. Learning and memorial ability of the rats were observed by moving-back with current stimulation experiment; expression of synaptohydin protein in hippocampus of exercise-fatigue rats treated with Sinisan for 10 days were detected by immunohistochemistry technique and compared to control group and model group.Results The moving-back time and the error times of the exercise-fatigue rats as follows: model group(73.00±61.96) s/(1.67±1.15), control group(144.25±57.14) s/(0.50±0.80)，Sinisan group (166.17±47.92) s/(0.38±0.49). Comparison of mean optical density of synaptohydin protein in hippocampus of rats in different groups as follows: control group (0.2636±0.10654) and Sinisan group (0.2555±0.163380) were higher than the model group (0.0474±0.1837)(P<0.05). The synaptohydin staining Results showed that synaptohydin-positive granules around the neuron in hippocampus of rats in control group were the darkest and densest, the Sinisan group were darker than the model group rats.Conclusion Sinisan could improve the learning and memorial ability and increase the synaptohydin protein in the hippocampus of the exercise-fatigue rats.