FIELD OF THE INVENTION

This invention pertains to obtaining cellular and genetic material from female reproductive system for various purposes including, without limitation, cytological analysis, sequencing, and detection of mutations for, without limitation, detecting, screening, and monitoring ovarian, fallopian tubal, or endometrial cancer or similar diseases.

BACKGROUND

Since the introduction of the Papanicolaou (“Pap”) test in 1950, the mortality of cervical cancer in screened populations has been reduced by more than 75%. In contrast, deaths from other cancers of the female reproductive system—cancers of ovaries, fallopian tubes, and uterus—have not substantially decreased during that same time period. As a result, more than 60,000 women in the U.S. are diagnosed with ovarian, fallopian tubal, and endometrial (uterus lining) cancer each year. Although endometrial cancer is more common than ovarian cancer, the latter is more lethal. In the U.S., approximately 15,000 and 8,000 women are expected to die each year from ovarian and endometrial cancers, respectively. World-wide, over 200,000 deaths from these tumors are expected this year alone. The mortality associated with undetected gynecologic malignancies has made the development of effective screening tools and methods a high priority.

The Pap test is currently provided to tens of millions of women in tens of millions medical offices every year, usually during a routine gynecological examination. The procedure is non-invasive, painless, and fast. It involves inserting a brush (an endocervical brush) into the cervix and rotating the brush inside the cervix to collect cervical cells by gently scraping the cervical canal. Then the brush with the collected material is rinsed in a vial to release the cervical cells and discarded. The collected cells released into the rinsing liquid are analyzed for abnormalities indicative of cervical cancer and the DNA is analyzed for the presence of HPV, the causative agent of cervical cancer.

As the cervical canal is usually filled with cervical mucus, the material collected on the brush contains some visible amount of this mucus. This mucus within each sample normally has varying viscosity and solubility—from almost liquid consistency to almost gelatinous. After the brush with the sample is rinsed, the cervical mucus partly ends up released into the rinsing liquid and partly remains on the brush and ends up discarded with it.

SUMMARY

As described above, the cervical mucus remaining on the brush after rinsing is discarded in the course of the Pap test. Also, during the rinsing, some cervical mucus falls off or detaches from the brush without dissolving in the rinsing liquid; such undissolved cervical mucus also remains unused during Pap tests.

This undissolved cervical mucus on the brush and/or at the bottom of the rinsing vial, hereinafter called “rinsed cervical mucus” or just “rinsed mucus”, is the focus of the following example embodiments.

Efficient harvesting of cells, cell fragments, free nuclei, and DNA material (“DNA material”, as used in this application, includes, without limitation, DNA, DNA fragments, genomic DNA, and/or cell-free DNA) from female reproductive system is made possible by processing the rinsed cervical mucus (usually, gelatinous part of cervical mucus samples).

These cells, cell fragments, free nuclei, and/or DNA material may originate, for example, from ovarian, fallopian tubal, and endometrial tumors, benign or cancerous.

This DNA material accumulated in the rinsed cervical mucus can be harvested from the rinsed cervical mucus for a variety of medical or biological purposes.

If someone would choose to run a genomic analysis on the DNA material obtained from Pap test rinsing liquid, any information about ovarian, fallopian tubal, and endometrial tumors, if present, would be drowned by the overwhelming amount of genetic information from cervical cells.

On the other hand, the number of cervical cells present in the rinsed cervical mucus used in the example embodiments is significantly lower in comparison, making the DNA material harvested according to the disclosed methods much more valuable for genetic, genomic and epigenetic analysis.

This DNA material accumulated in the rinsed cervical mucus may be released from the mucus into a solution by a variety of methods including, without limitation, mechanical vibrations and other types of mechanical impact, by treating the mucus with emulsifier, or other chemical treatment to dissolve or reswell the rinsed cervical mucus.

The embodiments include a method comprising rinsing a cervical mucus sample comprising cervical cells in a rinsing liquid to release the cervical cells into the rinsing liquid and to produce a rinsed mucus sample; separating the rinsed mucus sample from the rinsing liquid and the cervical cells released into the rinsing liquid; and releasing of cells, cell fragments, free nuclei, and/or DNA material from the rinsed mucus sample.

Some embodiments further comprise lysing the cells, cell nuclei, or cell fragments released from the rinsed mucus sample.

In some embodiments, the releasing of cells, cell fragments, free nuclei, and/or DNA material from the rinsed mucus sample and the lysing of the released cells, cell nuclei, or cell fragments released from the rinsed mucus sample are performed simultaneously.

In some embodiments, the number of cervical epithelial cells rinsed off the cervical mucus sample during the rinsing of the cervical mucus sample in the rinsing liquid is greater than the number of cervical epithelial cells released from the rinsed mucus sample during the releasing of cells, cell fragments, free nuclei, and/or DNA material from the rinsed mucus sample.

In some embodiments, the separating of the rinsed mucus sample from the rinsing liquid and the cervical cells released into the rinsing liquid comprises aspiration, filtration, or centrifugation.

In some embodiments, the rinsed mucus sample is located on a mucus collector after the rinsing of the cervical mucus sample in the rinsing liquid to produce the rinsed mucus sample.

In some embodiments, the separating of the rinsed mucus sample from the rinsing liquid and the cervical cells released into the rinsing liquid comprises removing the mucus collector from the rinsing liquid.

In some embodiments, the cervical mucus sample is located on the mucus collector before the rinsing of the cervical mucus sample in the rinsing liquid to release the cervical cells into the rinsing liquid and to produce the rinsed mucus sample.

In some embodiments, the releasing of cells, cell fragments, free nuclei, and/or DNA material from the rinsed mucus sample comprises a chemical or mechanical treatment of the mucus sample or a combination thereof or treating the rinsed mucus sample with a compound comprising an emulsifier.

In some embodiments, the releasing of cells, cell fragments, free nuclei, and/or DNA material from the rinsed mucus sample comprises at least partially dissolving or reswelling the rinsed mucus sample in an emulsifier solution.

Some embodiments further comprise removing cells from the emulsifier solution and isolating DNA material from the emulsifier solution.

In some embodiments, the removing of cells from the emulsifier solution comprises filtration or centrifugation.

Some embodiments further comprise removing mucus from the emulsifier solution before isolating DNA material from the emulsifier solution.

The embodiments also include a method comprising rinsing a cervical mucus sample comprising cervical cells in a rinsing liquid to release the cervical cells into the rinsing liquid and to produce a rinsed mucus sample; separating the rinsed mucus sample from the rinsing liquid and the cervical cells released into the rinsing liquid; and lysing of cells, cell nuclei or cell fragments contained in the rinsed mucus sample.

In some embodiments the lysing of cells, cell nuclei, or cell fragments contained in the rinsed mucus sample comprises lysing the rinsed mucus sample.

The embodiments further include a method comprising rinsing a cervical mucus sample comprising cervical cells in a rinsing liquid to release the cervical cells into the rinsing liquid; removing the cervical cells released into the rinsing liquid from the rinsing liquid; and isolating DNA material from the rinsing liquid.

In some embodiments, the removal of cervical cells released into the rinsing liquid from the rinsing liquid comprises filtration or centrifugation.

Some embodiments further comprise removing mucus from the rinsing liquid before isolating DNA material from the rinsing liquid.

Some embodiments further comprise at least partially dissolving or reswelling the cervical mucus sample before the removal of the cervical cells released into the rinsing liquid from the rinsing liquid.

In some embodiments, the at least partially dissolving or reswelling of the cervical mucus sample before the removal of the cervical cells released into the rinsing liquid from the rinsing liquid occurs in a solution comprising emulsifier.

The embodiments further include a method comprising lysing a cervical mucus sample and cells contained therein in a lysing liquid to lyse the cervical mucus sample to release DNA material from the cervical mucus sample into the lysing liquid; and isolating DNA material from the lysing liquid.

