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Method for screening cervical cancer

A technology of cervical cancer and screening method, which is applied in the field of cervical cancer diagnostic reagents, can solve problems such as the performance cannot meet the needs, the type of cervical cancer cannot be judged, and the information of the type of cancer cannot be obtained, so as to achieve high precision and high efficiency effect

Inactive Publication Date: 2005-10-05
SYSMEX CORP
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  • Abstract
  • Description
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Problems solved by technology

[0009] However, since the markers used in U.S. Patent Application Publication No. 2002 / 0106685 are markers of general cancer cells, even if the presence or absence of cancer cells can be determined, the type of cervical cancer cannot be determined.
The marker disclosed in U.S. Patent No. 5,858,683 is a marker specific to cervical cancer, but since it is a common marker for squamous cell carcinoma and adenocarcinoma, it is still not possible to obtain information for distinguishing the type of cancer
[0010] In addition, it is reported that the NMP179 antibody can react with some normal cells (ActaCytologica, Volume 43, Number 6 / November-December 1999: 1015-1021), but the performance cannot meet the needs

Method used

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preparation example Construction

[0090] [Preparation of samples for measurement]

[0091] The assay samples provided for the screening method of the present invention were prepared as follows.

[0092] First, the cervical cancer diagnostic reagent of the present invention containing the first labeled antibody, the second labeled antibody, and the third labeled antibody is added to cervical cells to be screened, and reacted for a predetermined time to prepare a measurement sample.

[0093] Cervix cells may be aggregates of cells dispersed into single cells, or cells on a smear specimen. In order to be applicable to a flow cytometer with a high processing speed, it is preferable to use aggregates of cells dispersed into single cells.

[0094] The cervix cell group that is used for measuring the sample preparation can directly use the cervix cell mass that is taken when wiping the surface of the cervix with a cotton swab or a scraper, or it can be mixed with the cervix of the present invention after removing mu...

Embodiment 1

[0172] Prepare 4 test tubes, add approximately 1×10 6 Individual HeLa cells, C33A cells, clinical samples 1, 2. After the cells were fixed with PreservCyt solution from Cytyc Company, they were centrifuged at 10,000 rpm for 1 minute, and the supernatant was discarded. Then 5% N-acetyl-L-cysteine ​​was added, stirred, centrifuged at 10000 rpm for 1 minute, and the supernatant was discarded. After the cells were washed with PBS, they were centrifuged at 10,000 rpm for 1 minute, and the supernatant was discarded. 1 ml of PBS or PBS-T (PBS containing 0.05% Tween20) containing 1% goat serum was added, stirred, and then blocked for 10 minutes.

[0173] After blocking, centrifuge at 10,000 rpm for 1 minute, and discard the supernatant. The reagent containing the first labeled antibody prepared above was added, and the reaction was stirred at room temperature for 30 minutes.

[0174] After the antibody reaction, centrifuge at 10,000rpm for 1 minute, discard the supernatant, wash t...

Embodiment 2

[0185] Add about 1×10 oral mucosal epithelial cells to the test tube 6 Each, in order to prevent non-specific staining caused by RNA, 100 μl of 300 μg / ml RNase A (Sigma #R-4612) diluted in PBS was added. After stirring at room temperature for 5 minutes, centrifuge at 10,000 rpm for 1 minute, and discard the supernatant. Then, 500 µl of 10 µM PI (Propidium iodide) was added as a solution for nuclear staining, and stirred at room temperature for 30 minutes. Centrifuge at 10,000 rpm for 1 minute, discard the supernatant, add 500 μl of 0.05% PBST, and centrifuge at 10,000 rpm for 1 minute. After replacing the solution with an appropriate amount of PBS, add to the Figure 6 Flow cytometers constructed as indicated.

[0186] Excited with an argon ion laser at 488 nm, while recording the distribution of orange fluorescence from nuclear staining and the distribution of side scattered light, an image of cells passing through the flow cell was created. Figure 22 The nuclear staining ...

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Abstract

The invention provides a reagent for diagnosing cervical cancer, which can not only detect the presence or absence of cervical cancer but can also distinguish squamous cell carcinoma and adenocarcinoma from each other, a method for screening cervical cancer by using the reagent, particularly a screening method capable high speed processing by utilizing flow cytometry. The reagent comprises a first labeled antibody reacting with gland cells, a second labeled antibody reacting with adenocarcinoma cells and a third labeled antibody reacting with atypical cervical squamous cells, the antibodies being labeled with mutually distinguishable labels respectively. Preferably, at least one selected from the group consisting of MUC1 antibody, cytokeratin 7 antibody, and cytokeratin 18 antibody is used as the first labeled antibody, at least one selected from the group consisting of cytokeratin 8 antibody and HIK1083 antibody is used as the second labeled antibody, and at least one member selected from the group consisting of NMP179 antibody, p16[INK4A] antibody, Ki-67 antibody, p53 antibody, p21 antibody, EMA antibody, CEA antibody and MIB-1 antibody is used as the third labeled antibody.

Description

technical field [0001] The present invention relates to a cervical cancer diagnostic reagent capable of detecting the presence or absence of squamous cell carcinoma and its precancerous state, adenocarcinoma and its precancerous state, and cervical cancer using the diagnostic reagent for a sample of cervical cell group collected from a living body A screening method for cancer, and an automatic diagnosis method for automatic determination of adenocarcinoma and squamous cell carcinoma. Background technique [0002] As a screening method for early detection of cervical cancer, cell diagnosis can be effectively used in medical diagnosis and the like. [0003] Among them, the cell diagnosis of cervical cancer can be made by rubbing the surface of the cervix with a cotton swab or scraper, and then directly applying the scraped cells on a glass slide to make a specimen, and then observing it with a microscope. The reality of diagnosis by observing cell morphology with a microscop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/58
CPCG01N33/57411G01N33/582
Inventor 中野浩一石坂正树岸和希渡边美晴井邨泰之
Owner SYSMEX CORP
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