Method for detecting bacterium serotypes
A technology of serotype and bacteria, applied in the direction of biochemical equipment and methods, measurement/testing of microorganisms, resistance to vector-borne diseases, etc., can solve problems such as high cost and complexity
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Embodiment 1O157
[0312] The detection of embodiment 1.O157 serotype Escherichia coli
[0313] In this example, the reagents described in Table 1 (9 detection probes, 38 mediator probes, 38 upstream primers, 38 downstream primers, and 1 universal primer) and the detection protocol described in Table 2 were used , to detect samples containing Escherichia coli O157 serotype.
[0314] In short, in this example, a 25 μL PCR reaction system was used for real-time PCR, and the PCR reaction system included: 1×PCR buffer (67mM Tris-HCl, 50mM KCl and 0.085mg / mL BSA), 6.0mM MgCl 2 , 0.2 mM dNTPs, 2.0 U of polymerase TaqHS (Takara), various reagents described in Table 1 (used at the indicated working concentrations), and 5 μL of E. coli DNA of serotype O157. The reaction conditions of real-time PCR were: 95°C, 5min; then 50 cycles (95°C, 20s and 63°C, 1min). After PCR is finished, carry out melting curve analysis according to following program: 95 ℃, 2min; The temperature was raised from 40°C to 95°C, ...
Embodiment 2 38
[0316] Embodiment 2. Thirty-eight multiple detections
[0317] In this example, the reagents described in Table 1 (9 detection probes, 38 mediator probes, 38 upstream primers, 38 downstream primers, and 1 universal primer) and the detection protocol described in Table 2 were used , to contain 37 kinds of bacterial serotypes (respectively from O29 type E. coli, O29 type E. coli, O26 type E. coli, O169 type E. coli, O161 type E. , E. coli O1, E. coli O157, E. coli O148, E. coli O27, E. coli O167, E. coli O6, E. coli Gp6, E. coli O15, E. coli O159, E. coli Gp12 , E. coli O25, E. coli O166, E. coli O91, E. coli Gp2, E. coli O86, E. coli O111, E. coli O121, E. coli O145, E. coli O55, E. coli Gp9 , E. coli O63, E. coli O18, E. coli O128, E. coli O119, E. coli O153, E. coli O103, E. coli O113, E. coli O104, E. coli O45, E. coli O142 DNA) and control DNA (Escherichia coli yccT gene) samples were tested.
[0318] In short, in this example, a 25 μL PCR reaction system was used for re...
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