After the clinic has evaluated the couple for several male and female fertility factors, in addition to the woman’s age, they determine the most appropriate treatment option for the couple. One of the determining factors for egg quality is the Day-3 FSH value. A high FSH predicts poor egg quality. This determines the treatment protocol (GnRH agonist or antagonist protocol).

Once a particular treatment protocol has been determined there are 5 different stages of the IVF procedure.

1. Ovarian stimulation: The goal in this stage is to obtain at least 8-10 mature eggs. The stimulation cycle consists of 8 days starting from Day 2 of the menstrual cycle. Blood work and ultrasound are done 3-4 times during this period. The blood work usually consists of E2 and FSH. There is a direct correlation between the E2 value and the number of eggs/follicles. The physician uses the E2 and ultrasound result to determine if the treatment is going well. If a patient is hyper-stimulated and develops too many eggs, the cycle is usually cancelled. It is therefore very important that the physician be able to review the E2 result and ultrasound on the same visit. Tosoh’s ST AIA-PACK E2 offers results in 18 to 20 minutes. If the blood is drawn as soon as the patient checks-in, the E2 result will be available to the clinician during the ultrasound.

2. hCG trigger: An hCG injection is given on the 9th or 10th day to induce ovulation and final egg maturation. This depends on the size of a mature follicle. A follicle is considered mature if the diameter is approximately 18-20 mm. The E2 level at this stage is around 1000-4000 pg/mL. On the day of the HCG trigger, the E2/follicle ratio should be approximately 100-200 and the progesterone level should be 0.8 ng/mL. Since E2 levels are very high, it is desirable to have an assay that offers a broad assay range. Tosoh’s ST AIA-PACK E2 offers an assay range of 25 to 3000 pg/mL.

3. Egg retrieval: The egg retrieval is planned for 34-35 hours after hCG injection and shortly before that time, ovulation may begin. At this stage the patient should produce enough progesterone for a successful implantation. Progesterone supplements are usually given to prepare the uterus for the implantation.

4. Embryo transfer: After retrieval, the egg is fertilized with the partner’s sperm. The embryo can be transferred on day 3 of fertilization or day 5 (blastocyst) of fertilization. The embryo is carefully placed into the patient’s uterus. Usually 2-3 embryos are transferred and vary from patient to patient.

5. Beta hCG test: If the implantation is successful, the beta hCG test will be positive on the 14th day after egg retrieval. A positive beta hCG indicates pregnancy. The beta hCG doubles very 48 hours and is monitored throughout the pregnancy. The patient usually begins routine pre-natal care by her obstetrician at 12 weeks of pregnancy.

IUI:
In an IUI procedure the patient goes through stages 1 and 2. The stimulation protocol is usually mild to get 1-2 mature follicles. Thirty-four to thirty-five hours after the hCG trigger, the patient is inseminated with the partner’s sperm. The fertilization occurs naturally.

Egg Donation:
If an egg donor is used, the donor goes through stages 1 to 3. Once the egg is retrieved, it is fertilized with the partner’s sperm. The recipient goes through all the stages but the embryo transferred utilizes the egg from the donor.

The IVF process is very involved and has an emotional impact on both the patient and the care givers. Because it is very time sensitive, an assay with fast turnaround time is always desirable. Operators using immunoassay analyzers have to multitask, as both embryologist and andrologist in many clinics. A simple, easy to operate instrument is a good choice. The most important criterion for immunoassays in IVF clinics is the reliability of the assays. An E2 assay with low CVs is important as the day-to-day change in result is monitored throughout the stimulation cycle.

Tosoh offers a very good solution with its menu of reproductive assays. The assay time is 18-20 minutes, which allows the physician time to review the results with the patient on the same visit. Tosoh minimizes heterophile antibody interference by using special Ab coating methods and HAMA blockers in the assays. CVs on the E2 are <8.9%. 90 day calibration stability allows the laboratory to test throughout the cycle, with no re-calibration. All of these features of Tosoh’s reproductive assays ensure consistent, reliable results.

Tosoh’s extensive immunoassay test menu provides a wide range of immunoassay tests including thyroid, cardiac markers, tumor markers, reproductive hormones, anemia markers, metabolic markers and more. All Tosoh AIA-PACK assays now carry 90 day calibration curve stability with the exception of Troponin I, B12 and Folate, which will continue to have the 30 day calibration stability.

AIA-PACK test cups and trays are labeled with code and lot number for automated scheduling and inventory management. A 10 minute incubation protocol provides fast turnaround time. Additionally Tosoh AIA reagents have a shelf life of 1 year from the date of manufacture.

Because AIA-PACK test cup reagents are interchangeable on all Tosoh AIA systems, inventory management is easy and problem free. Transition from one system to another is seamless, ensuring consistent results and efficient, economical operation for the laboratory.

Tosoh Bioscience, Inc. (TBI) provides highly sophisticated diagnostic systems for immunoassay and HPLC testing to doctor’s offices, hospitals and reference laboratories throughout the Americas. Based in South San Francisco, CA, TBI is a U.S. subsidiary of Tosoh Corporation’s Bioscience Division, headquartered in Tokyo, Japan.

Testimonial:
The University of Virginia Medical Center runs 600 samples/year, and uses the G7 Beta-thalassemia mode because it is faster than electrophoresis: “The Beta- thalassemia program on the G7 has reduced my turnaround time from 7 days to 24 hours”- Walter Oliveira.

Hemoglobinopathies may interfere with an HbA1c result which merits further investigation. Please contact your local System Sales Specialist to learn how you can help provide better patient care.

Tosoh is investigating the clinical utility of a real-time, nucleic acid-based assay for CMV monitoring, called the TRC-CMV assay. If you would like more information, please call 1-800-695-6550, ext. 8117.

Calibrations should be performed:
- With each new lot number of AIA-PACK Test Cups.
- After the defined calibration period has expired.
- As QC dictates
- When major service has been completed and/or component changed that could affect calibration.
- Remember to review all flags

Tosoh Technical Support

Dr. John Murphy, Senior TBI Manager Technical Support

It is important to review and question any chromatogram with the following characteristics:
- The SA1c is outside the normal range established by your laboratory.
- The SA1c peak is not detected
- Total Area is outside of the range for your Instrument
- An unidentifiable peak (e.g. P00) appears before the A0 peak

Tosoh Technical Support

Dr. John Murphy, Senior TBI Manager Technical Support

RBC folate levels are a more accurate reflection of body stores of folate. Although serum folate offers a reliable test, it fluctuates based on dietary changes. RBC folate concentration correlates well with other indexes of folate deficiency such as bone-marrow cell morphology and hematological changes.

Tosoh’s AIA-PACK RBC Folate offers an assay range of 0.62 to 24 ng/mL. The assay will be available in the 40 minute incubation format on all AIA platforms (with the exception of AIA-360). To order: call 1-866-527-3587, using Catalog #: 020214.

References:

http://www.labtestsonline.org/understanding/analytes/vitamin_b12/test.html

AIA-PACK RBC Folate – AIA Analyte Application Manual.
Tosoh Bioscience, Inc., South San Francisco, CA.
(September 2009)

A: Call the toll free number 800 24-TOSOH (800-248-6764). Normal business hours are Mon-Fri 6 AM – 5 PM Pacific Daylight Time. On weekends and after normal business hours, these calls are routed through an answering service, and the call is returned.

Q: Who do I call for periodic maintenance?

A: All requests for service or periodic maintenance should be placed with Tosoh Technical Support. To expedite the process, it is always helpful to have your customer number and instrument serial number available when you call.

Q: What number do I call for placing orders or checking on the status of orders?

A: To place an order or for questions about orders, please direct these calls to Customer Service (Order Processing) at 866-527-3587 or fax your order to 800-685-7595. If you do not buy directly from Tosoh, please place your order through your preferred distributor.

Our website www.tosohbioscience.us has all of the contact information under the Contact Us link at the top of the page.

Dr. John Murphy, Senior TBI Manager Technical Support

Analyte independent interferences refer to common interferences like hemolysis, lipemia, effects of anti-coagulant and sample storage. The term pre-analytical variables are commonly used for these types of interferences.

Analyte dependent interferences refer to interaction between constituents in the sample with one or more reagent antibodies. They include compounds with chemical differences but structural similarities that cross react with the antibody i.e. heterophile antibodies, human anti-mouse antibodies (HAMA), rheumatoid factors and other proteins.

Interferences can cause false positive and false negative results. RF, HAMA and heterophile antibodies bind to the Fc portion of the IgG molecule and cause false positive results (Fig A). The interferences can bind to the capture Ab due to structural similarities and cause false negative results (Fig. B).

Immunoassay manufacturers take several steps to minimize interference (Fig. C).
1. Characterize epitypes to increase Ab specificity.
2. Use antibody coating methods that minimize the exposure of the Fc portion of the IgG.
3. HAMA blockers.

It is sometimes necessary for laboratories to confirm the presence of HAMA or other interfering substances and pre-treat the sample to neutralize interference.