Method of assessing colorectal cancer by measuring hemoglobin and M2-PK in a stool sample
A colorectal cancer, hemoglobin technology, applied in the field of helping to evaluate colorectal cancer, can solve the problem of limited sensitivity of colon cancer
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Embodiment 1
[0101] research group
[0102] Stool samples from 94 well-characterized CRC patients using the UICC classification given in Table 1 were used.
[0103] Table 1: CRC samples and corresponding UICC classifications
[0104] Phases according to UICC
Number of samples
UICC 0
2
UICC I
21
UICC II
27
UICC III
36
UICC I-III (non-IV, categories I to III are not individually phased)
8
Total number of CRCs
94
[0105] The CRC samples of Table 1 have been evaluated compared to control samples obtained from individuals who underwent colonoscopy and did not have adenomas, polyps, or colorectal cancer. Table 2 gives an overview of the controls used:
[0106] Types of control patients
Embodiment 2
[0108] Extraction of stool samples for determination of hemoglobin and M2-PK
[0109] For each patient studied, two stool aliquots were collected at different locations on a single bolus prior to colonoscopy or surgery. A total of about 1 gram per stool sample was collected using specific collection equipment from Sarstedt (Id. no. 80.623.022). Fecal samples thus collected were frozen as soon as possible and stored at -70°C until extraction.
[0110] For the determination of hemoglobin, 100 mg of stool samples were placed in 2 mL Eppendorf cups for each extraction experiment. The 100 mg stool samples were extracted by using 1 ml fresh extraction buffer.
[0111] Use the following extraction buffers:
[0112] 9,49g Na 2 HPO 4 ×2H 2 o
[0113] 1,84g KH 2 PO 4
[0114] 1g NaN 3
[0115] 0,4g Na 2 EDTA×2H 2 o
[0116] 10ml chicken albumin
[0117] 50ml Nonidet P40 10% w / v
[0118] 1 tablet Complete mini (Roche Diagnostics, Id.No.1836145) was added to 1 L with doub...
Embodiment 3
[0122] Immunoassay for Determination of Hemoglobin and M2-PK from Extracts of Fecal Samples
[0123] 3.1 Hemoglobin
[0124] Hemoglobin determination was performed using the "Haem Immun" assay (Labor Limbach, Heidelberg) according to the manufacturer's instructions. 10 [mu]l of Hb extract was used as sample in the immunoassay.
[0125] 3.2M2-PK
[0126] The determination of M2-PK was used according to the instructions given by the manufacturer " Tumor M2-PK" assay (Schebo Biotech AG, Giessen). 50 μl of M2-PK extract was used as sample in this immunoassay.
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