IPC분류정보
국가/구분 |
United States(US) Patent
등록
|
국제특허분류(IPC7판) |
|
출원번호 |
US-0320558
(2010-05-17)
|
등록번호 |
US-9182403
(2015-11-10)
|
국제출원번호 |
PCT/IL2010/000399
(2010-05-17)
|
§371/§102 date |
20111115
(20111115)
|
국제공개번호 |
WO2010/134073
(2010-11-25)
|
발명자
/ 주소 |
- Idelevich, Pavel
- Elkeles, Adi
- Terkieltaub, Dov
- Eyal, Ami
|
출원인 / 주소 |
|
대리인 / 주소 |
|
인용정보 |
피인용 횟수 :
0 인용 특허 :
20 |
초록
▼
Provided are methods and kits for staining cervical cell sample by contacting the cervical cell sample with a Ficus elastics plant extract, staining the cervical cell sample with New Fuchsin, and staining the cervical cell sample with Light Green or Fast Green. Also provided are method of diagnosing
Provided are methods and kits for staining cervical cell sample by contacting the cervical cell sample with a Ficus elastics plant extract, staining the cervical cell sample with New Fuchsin, and staining the cervical cell sample with Light Green or Fast Green. Also provided are method of diagnosing a pre-malignant or a malignant cervical tumor in a subject, by staining the cervical cell sample and identifying at least one cervical cell having a red cytoplasm above a pre-determined threshold, wherein presence of the at least one cervical cell having the red cytoplasm above the pre-determined threshold is indicative of a non- or less-differentiated cell as compared to a normal cervical cell, thereby diagnosing the pre-malignant or a malignant cervical tumor in the subject.
대표청구항
▼
1. A method for differential staining a liquid-based cervical monolayer cell sample, the method comprising the steps of: (i) incubating a liquid-based cervical monolayer cell sample devoid of blood cells with trichloroacetic acid;(ii) contacting the cervical cell sample with a Ficus elastics plant e
1. A method for differential staining a liquid-based cervical monolayer cell sample, the method comprising the steps of: (i) incubating a liquid-based cervical monolayer cell sample devoid of blood cells with trichloroacetic acid;(ii) contacting the cervical cell sample with a Ficus elastics plant extract,(iii) staining the cervical cell sample with new fuchsin,(iv) staining the cervical cell sample with light green or fast green, and(v) staining the cervical cell sample with an agent which differentially stains mucin in the cervical cell sample in a color distinguishable from the color obtained by staining with new fuchsin, light green and fast green to differentially stain the cervical cell sample. 2. A method of diagnosing a pre-malignant or a malignant cervical tumor in a subject, the method comprising the steps of: (a) staining a liquid-based cervical monolayer cell sample of the subject according to the method of claim 1, to obtain a stained cervical cell sample,(b) identifying at least one cervical cell of said cervical cell sample having a red cytoplasm above a predetermined threshold, wherein a presence of said at least one cervical cell having said red cytoplasm above said predetermined threshold is indicative of a non- or less differentiated cell as compared to a normal cervical cell, thereby diagnosing the pre-malignant or the malignant cervical tumor in the subject. 3. The method of claim 2, further comprising the step of detecting an expression level of a cervical malignant marker or a cervical pre-malignant marker in at least one cervical cell of the cervical cell sample, wherein an expression level above a pre-determined threshold is indicative that said at least one cervical cell is a malignant or a pre-malignant cell, respectively, thereby diagnosing the pre-malignant or the malignant cervical tumor in the subject. 4. The method of claim 3, wherein said cervical malignant or cervical pre-malignant marker is selected from the group consisting of: ki67 antigen (antigen identified by monoclonal antibody Ki-67), Mtn2, Hypoxiainducible factor 1-alpha, Id-1, p161 NK 4a, p21 WAF1/CI P1, Tn antigen, P53 and proliferating cell nuclear antigen. 5. The method of claim 3, further comprising the step of: (c) analyzing a morphology of said at least one cervical cell having said red cytoplasm above said pre-determined threshold, wherein a presence of an abnormal morphology in said same cell having said red cytoplasm above said pre-determined threshold as compared to a morphology of a normal cervical cell sample indicates the positive diagnosis of the pre-malignant or the malignant cervical tumor in the subject. 6. The method of claim 1, further comprising the step of: (vi) staining the cervical cell sample with hematoxylin. 7. The method of claim 1, further comprising the step of digesting the mucin in the cervical cell sample with an enzyme prior to said staining with new fuchsin. 8. The method of claim 1, further comprising the step of mounting the stained cervical cell sample with amounting medium comprising an anti-oxidant. 9. The method of claim 2, wherein said pre-malignant or malignant cervical tumor is selected from the group consisting of: an intraepithelial neoplasia, a squamous intraepithelial lesion, carcinoma in situ, invasive carcinoma, squamous cell carcinomas and adenocarcinoma. 10. The method of claim 6, wherein said step of staining said cervical cell sample with hematoxylin is effected prior to said step of contacting said cervical cell sample with Ficus elastics plant extract. 11. The method of claim 1, wherein said Ficus elastics plant extract is selected from the group consisting of: a crude Ficus elastics plant extract, a crude ethanol extract of the Ficus elastics plant, a crude ethanol extract of the Ficus elastics plant, an ethanol extract of leaf tissue of Ficus elastics plant and proanthocyanidins. 12. The method of claim 3, wherein said step of detecting the expression level of said cervical malignant or said cervical pre-malignant marker is performed using a high affinity agent selected from the group consisting of: an antibody which specifically binds to said cervical malignant or cervical pre-malignant marker and a polynucleotide. 13. The method of claim 12, wherein said polynucleotide is selected from the group consisting of a polynucleotide which specifically hybridizes to an RNA encoding said cervical malignant or cervical pre-malignant marker and a polynucleotide which specifically amplifies said cervical malignant or cervical pre-malignant marker.
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