Assay and kit for predicting implantation success in assisted fertilisation
The present invention relates to an assay for determining the implantation potential of a plurality of embryos each obtained or to be obtained by assisted fertilization of an oocyte of a female subject, comprising measuring the levels of granulocyte-colony stimulating factor (G-CSF) in the follicular fluid (FF) present in the follicle from which each oocyte is derived, and determining the implantation potential of each embryo from the level of measured FF G-CSF. It also relates to a kit for carrying out the assay. It further relates to a method for assisted fertilization.
1. An assay for determining for a female subject the implantation potential of embryos obtained or to be obtained by assisted fertilisation comprising:
(i) measuring, for a plurality of ooctyes collected from said subject, the level of follicular fluid granulocyte-colony stimulating factor (G-CSF) present in the follicular fluid (FF) of a follicle of each collected oocyte; and
(ii) determining from the levels of FF G-CSF measured, the implantation potentials of the embryos obtained or to be obtained by assisted fertilisation of the oocytes.
2. The assay according to claim 1 , wherein oocytes having the highest levels of FF G-CSF have the highest potential of implantation.
3. The assay according to claim 1 , wherein each sample of FF is obtained from a follicular aspirate.
4. The assay according to claim 1 , wherein respective levels of FF G-CSF are measured within 20 hours of collection of the follicular aspirate.
5. The assay according to claim 1 , wherein a level of FF G-CSF in a follicle which is equal to or less than 20.6 pg/ml determines no or a low potential of implantation.
6. The assay according to claim 1 , wherein a level of FF G-CSF in a follicle which is equal to or greater than 24.0 pg/ml determines a high potential of implantation.
7. The assay according to claim 1 , wherein the respective levels of FF G-CSF are measured using an immunoassay.
8. The assay according to claim 1 , wherein the respective levels of FF G-CSF are measured using a competitive or immunometric assay.
9. The assay according to claim 1 , wherein the respective levels of FF G-CSF are measured using a Luminex assay.
10. The assay according to claim 1 , wherein the respective levels of FF G-CSF are measured by any of surface plasmon resonance, fluorescence resonance energy transfer, bioluminescence resonance energy transfer, fluorescence quenching fluorescence, fluorescence polarization, mass spectrometry (MS), high performance liquid chromatography (HPLC), HPLC/MS, HPLC/MS/MS, capillary electrophoresis, rod or slab gel electrophoresis.
11. A method for assisted fertilization of a female subject comprising:
(i) collecting a plurality of oocytes from said subject,
(ii) determining the implantation potential for an embryo to be obtained from fertilization of each oocyte according to the assay of claim 1 ,
(iii) fertilizing the oocytes corresponding to embryos having a high potential for implantation, and
(iv) implanting the embryo so obtained into the female subject.
12. A method for assisted fertilization of a female subject comprising:
(i) collecting a plurality of oocytes from said subject,
(ii) determining the implantation potential for an embryo to be obtained from fertilization of each oocyte according to the assay of claim 1 ,
(iii) fertilizing the oocytes to obtain embryos, and
(iv) implanting the embryos having a high implantation potential.
13. The assay according to claim 1 , wherein the respective levels of FF G-CSF are measured using radio-immunoassay (RIA), enzyme-linked immunosorbent assay (ELISA) or enzyme-linked immunosorbent spot (ELISPOT) assay.