Method for synchronous detection of multiple tubercle bacillus specific secretion antigens
A technology for simultaneous detection of Mycobacterium tuberculosis, applied in the field of medical testing, can solve the problems of low sensitivity and specificity, and achieve the effect of improving sensitivity and improving the positive detection rate.
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Embodiment 1
[0030] 1) Through the analysis of the gene sequence of the antigen with strong specificity to Mycobacterium tuberculosis, several major secreted antigens of ESAT-6, MPT53, MPT64, MPT63 and MPT70 are determined as candidate genes.
[0031] Different fusion methods were used for vector construction and expression. The vector is pET21a, which contains 6×His at the C-terminus after expression, and pProEX-HTa, which contains 6×His at the N-terminus after expression. Table 1 shows the results of different fusion methods and vector construction and expression.
[0032] carrier
expression product
deduced product sequence
The expression level
pET21a
pET21a
pET21a
pProEX-HTa
pProEX-HTa
pProEX-HTa
MPT63-6×His
MPT70-6×His
MPT63-MPT70-6×His
6×His-MPT64
6×His-ESAT-6-MPT53
6×His-ESAT-6-MPT53-MPT64
SEQ1
SEQ2
SEQ3
SEQ4
SEQ5
SEQ6
Low expression level, unable to purify
Low expr...
Embodiment 2
[0054] Example 2 Serum Detection of Mycobacterium tuberculosis Infected Patients
[0055] With the step 6 method of embodiment 1, 30 parts of Mycobacterium tuberculosis infection patient's serum and 10 other case serums are tested, 24 routine test results of 30 routine TB positive patient's serums are judged as positive (80%), 10 routine TB negative serum samples all Negative (cutoff value 0.261).
[0056] Table 3 shows the test results of 30 tubercle bacillus infected patients' sera.
[0057] TB positive
serum sample
OD
value
TB positive
serum sample
OD
value
TB positive
serum sample
OD
value
TB negative
serum sample
OD
value
1
0.041
11
1.221
21
0.332
1
0.075
2
0.756
12
0.562
22
0.246
2
0.039
3
0.864
13
0.769
23
0.842
3
0.046
4
1.256 ...
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