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Diagnostic Testing: Infertility and Recurrent Miscarriages
The importance of Immune Pathological Evaluation of Pregnancies that Terminated in Spontaneous Miscarriage.
When a pregnancy fails, women often have a Dilatation and Curettage (D and C). When pregnancy tissue is obtained, it is sent to a pathologist usually associated with a laboratory or a hospital. Once the tissue is received, it is fixed in a formalin solution and embedded in paraffin (the final product is a paraffin block). The pathologist cuts the block into small slices, stains the tissue in the paraffin and looks at it under the microscope to determine that the tissue is pregnancy tissue and that it is normal and not infected. This is usually all that is done. Our Laboratory can do further testing (placental immunopathology) to determine if there is immune problem.
Immunology: Enhancing our Understanding What is Reproductive Immunology? History of Reproductive Immunology Diagnostic Testing: Infertility and Recurrent
Reproductive Failure: Causal Factors When to go for Immune Testing?
A. Recommended tests in Recurrent Pregnancy Loss:
Test Specimen Required
1. Chromosomal Analysis Tissue in Normal Saline
   a. POC- Product Of Conception
   b. Husband and Wife
Blood (4ml each in green- topped
2. Anti-Phospholipid Antibody
4ml Blood on red-topped vacutainer (Wife)
3. Reproductive Immunophenotype 8ml Blood in green-topped Vacutainer (Wife)
4. Cytokine Estimation
4ml Blood in green-topped vacutainer (Wife)
5. Estimation of Pregnancy Destroying Factor 4ml Blood in green -topped vacutainer (Wife)
6. Estimation of Mycobacterium tuberculosis 4ml Blood in green -topped vacutainer (Wife)
7. Immune Pathology Evaluation
   (a) Natural Killer Cells
   (b)Tumor Necrosis Factor
   (c) Pregnancy Destroying Factor
   (d) Macrophages
   (e) Progesterone Receptors

POC in 10% formalin or Paraffin Block
8. Screening for Infections Agents
   (a) Mycobacterium tuberculosis
   (b) Toxoplasma gondii
   (c) Cytomegalovirus
   (d) Herpes Simplex Virus
POC in 10% formalin or Paraffin Block
B. Recommended tests for IVF/IUI Implantation failure:
Test Specimen Required
1. Endometrial Receptivity Markers
   a. MAG
   b. Luteal-Phase Defect Markers
   c. Integrins

2. NK Cells
3. Tumor Necrosis Factor
4. Macrophages
5. Progesterone Receptors
6. Mycobacterium tuberculosis
Endometrium in 10% formalin


Paraffin Block
Recommended Test for Endometriotic Diseases
Test Specimen Required
1. Tumor Necrosis Factor (TNF-α)
2. Macrophages    
3. Progesterone Receptors 
4. Integrins (19 to 24th day)
5. Mycobacterium tuberculosis
Endometrium Biopsy


Endometrium Receptivity Test
Often couples have to overcome many hurdles before they can have a successful pregnancy. These include the considerable journey the sperm must make to reach the egg, the penetration of the egg by one successful sperm, the division of cells to create an embryo, the journey of the embryo to the uterus and, possibly the most difficult step of all, the attachment of the embryo to the uterine lining (implantation). Any abnormalities in the process of implantation are believed to be the basis of many cases of unexplained infertility in women. The question that researchers are trying to answer is: Can we help women for whom implantation has not been successful?

In many assisted reproductive technology (ART) programs, fewer than 10% of embryos successfully implant. Predictors of implantation success are needed, both to better understand the causes of infertility in women and to improve the efficacy and reliability of ART. Despite almost two decades of assisted reproductive technologies, the precise mediators of human implantation remain unknown. As a corollary to this problem, up to now no markers have existed to predict adequately whether implantation will occur during any given assisted cycle.
Uterine Receptivity Marker (Integrins):
Human endometrium undergoes a remarkable series of developmental changes during the menstrual cycle in preparation for embryonic implantation. The complex structure of the endometrium requires an array of distinct molecules which contribute to the cell distribution, adhesion, trafficking and signaling with matrix proteins of the endometrial meshwork. It has been documented, that these cellular activities are known to be mediated by a family of cell adhesion molecules termed as integrins (ITGs).

ITGs are cell surface glycoprotein receptors which consist of different -subunits and a common -sub-unit. So far 15  and eight  subunits have been identified on various cells. The involvement of ITGs in reproduction has been suggested recently. Several reasons make these molecules very attractive due to their constant involvement from egg to birth. The role of different ITG subunits in the assessment of endometrial maturation and receptivity has been reported. Lack or poor expression of some of the ITG subunits in endometrial cells leads to failure of embryo-endometrial interactions and implantation.
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