Compared with the rapid ELISA method, the microwave ELISA method has similar sensitivity and specificity to the rapid diagnosis of Schistosomiasis japonicum, and it is faster. The whole process only needs 5 minutes to develop color observation results, greatly improving the detection The working efficiency of schistosomiasis [1,2]. In this paper, two batches provided by the Institute of Parasitic Diseases of the Chinese Academy of Preventive Medicine were used to quickly observe the stability of the reagents under microwave conditions. The experimental results are reported below.
Compared with the rapid ELISA method, the microwave ELISA method has the similar sensitivity and specificity in the method of rapid diagnosis of Schistosomiasis japonicum by ELISA kit, and it has a faster speed. Improve the working efficiency of schistosomiasis detection [1,2]. In this paper, two batches provided by the Institute of Parasitic Diseases of the Chinese Academy of Preventive Medicine were used to quickly observe the stability of the reagents under microwave conditions. The experimental results are reported below.
1 Materials and methods
1.1 Kit
The rapid ELISA kits with batch numbers 200101 and 200305 provided by the Institute of Parasitic Diseases, Chinese Academy of Preventive Medicine were used.
1.2 Serum
In 2000, 15 sera from patients with acute schistosomiasis in Hubei Province (hereinafter referred to as acute blood) and 40 sera from patients with chronic schistosomiasis (hereinafter referred to as slow blood) were collected, a total of 55.
1.3 ELISA kit instrument
Galanz WP700L17 is a product of Shunde Galanz Electric Co., Ltd.
1.4 Method
In the ELISA kit experiment, 100μl of the test serum diluted 1:50 was dropped into the small hole of the antigen plate of the rapid ELISA kit, the household microwave oven was turned on, and the defrosting power was 340W according to Yuan Hong et al [2], and the radiation was 60s. After taking out, the reaction solution was poured out, rinsed with tap water three times, patted dry, and 100 μl of goat anti-human IgG was added dropwise, the same as the irradiation time and power. After taking out, the reaction solution was poured out as above, rinsed 5 times with tap water, patted dry, 2 drops of developer were added, and the reaction was observed. Each antigen plate is set with positive and negative standard serum controls.
1.5 Statistical processing chi-square test, anecdote and test.
2 results
2.1 Comparison of microwave ELISA and rapid ELISA The rapid ELISA kit with batch number 200101 was used, and the rapid ELISA kit with batch number 200305 was used as the control group. The enzyme conjugates were diluted in two experiments at 1: 1000 and 1: 500, respectively. The color was developed with the respective lotion reagents, and the experimental results were compared.
The results showed that when the enzyme conjugate was 1: 500 and 1: 1000, the specificity and sensitivity of the 200101 kit was not significantly different from the 200305 kit.
2.2 The color development of the positive reference and negative reference in the rapid ELISA kit with batch number 200305 under the condition of 1: 1000 enzyme conjugate is the control group, and the degree of color development is further observed and compared. The same color as the positive reference of the control group is recorded as (+), and the apparent color depth of the positive reference of the control group is (++), (+++), (++++), respectively. The negative reference is the same or lighter than the negative reference is marked as (-).
The results showed that when the same positive reference and negative reference were used as controls, the difference in positive reaction intensity between the 200101 kit and the 200305 kit was highly significant.
3 Discussion
The above results show that the two batches of kits 200101 and 200305 have significant differences in the experimental results. According to the instructions of the ELISA kit provided by the Institute of Parasitic Diseases of the Chinese Academy of Preventive Medicine, they should be stored in the refrigerator at 4 ℃ without opening , The validity period is 1 year, but when the storage conditions of the reagents are changed, the antigen reaction plate, enzyme conjugate, positive reference and negative reference are stored in a -25 ℃ refrigerator, the results show that low temperature (-25 ℃) still has good results to 2 years, The color-developing agent stored in a refrigerator at 4 ° C can still effectively develop color, which has certain significance for the preservation of the kit, and can be suitable for use in areas with few schistosomiasis patients. The same reference was made with the 200305 batch number, the positive reference and the negative reference of the enzyme conjugate 1: 500. The 200305 kit was significantly darker than the 200101 batch kit, and the difference in reaction intensity between the two batch kits was very significant. When the concentration of the enzyme conjugate is 1: 500, the color development is deeper than 1: 1000. It is concluded through experiments that for fast ELISA kits with a storage period of more than 1 year, it is recommended that the concentration of enzyme conjugate be 1: 500.
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