Luteinizing hormone (LH)

Luteinizing Hormone (LH) Detection Kit

Instructions for Use (Magnetic Microparticle Chemiluminescence Method)

Instruction of Luteinizing Hormone（LH）Detection Kit（Magnetic Solid Phase Chemiluminescent Immunoassay）

【Product Name】

Generic Name: Luteinizing Hormone (LH) Detection Kit (Magnetic Microparticle Chemiluminescence Method)

English Name: Luteinizing Hormone (LH) Detection Kit (Magnetic Solid Phase Chemiluminescent Immunoassay)

【Packaging Specifications】

50 tests/box, 100 tests/box, 200 tests/box.

Calibrator: 6 × 0.55mL (optional), 2 × 0.55mL (optional)

Quality Control: 2 × 0.75mL (optional)

【Intended Use】

The Luteinizing Hormone (LH) Detection Kit (Magnetic Microparticle Chemiluminescence Method) (hereinafter referred to as this kit) is used for the in-vitro quantitative detection of luteinizing hormone (LH) in human serum.

LH plays a very important role in the development and maturation of follicles and the regulation of the natural menstrual cycle. Low levels of LH in the follicular phase can hinder follicle development and reduce estrogen (E2) levels. The absence of an LH surge before ovulation can cause ovulation disorders, and low LH in the luteal phase can affect luteal function, even leading to luteal insufficiency. Luteal insufficiency refers to incomplete corpus luteum formation after ovarian ovulation, resulting in insufficient progesterone secretion, which prevents timely conversion of the endometrium, affecting endometrial receptivity and hindering embryo implantation, often leading to infertility or habitual abortion. The most common cause is insufficient LH surge secretion during follicle maturation. Abnormal LH levels and the resulting increase or decrease in follicle-stimulating hormone, estrogen, progesterone, and testosterone levels are associated with many diseases. Elevated LH levels in women can cause menopause, primary ovarian insufficiency, and polycystic ovary disease, while in men, it can cause primary gonadal insufficiency. Conversely, decreased LH levels in women can cause primary ovarian hyperfunction, and in men, it can cause primary gonadal hyperfunction. Therefore, detecting LH levels in the body is of great significance for disease diagnosis, prevention, and treatment. Common laboratory diagnostic methods in clinical practice include immunochromatography and chemiluminescence methods.

【Testing Principle】

This kit uses a direct sandwich method: biotin-labeled LH antibody, acridinium ester (AE)-labeled LH antibody, and LH in the sample, calibrator, or quality control combine to form a "sandwich" complex. Subsequently, streptavidin-coated magnetic microparticles are added. Through the specific binding of streptavidin and biotin, the antigen-antibody complex is connected to the magnetic particles. Under the action of an external magnetic field, the complex formed by the immunoreaction is separated from other unbound substances. After washing the complex, a pre-excitation solution and an excitation solution are added. Acridinium ester forms an unstable excited intermediate under the action of the pre-excitation solution and excitation solution. When the excited intermediate returns to the ground state, it emits photons, forming a chemiluminescent reaction, which can be detected using a chemiluminescence instrument. Within the detection range, the chemiluminescence intensity is proportional to the LH concentration in the sample. A modified four-parameter Logistic equation can be used to calculate the LH concentration in the sample.

【Main Components】

1. Product Composition

LH magnetic microparticle reagent: 1 bottle, streptavidin-coated magnetic microparticles;

LH antibody reagent 1: 1 bottle, biotin-labeled LH paired antibody conjugate;

LH antibody reagent 2: 1 bottle, acridinium ester (AE)-labeled LH antibody conjugate;

Calibrator (optional): 6 bottles or 2 bottles, containing BSA buffer with different amounts of LH antigen added. Calibrator values are obtained by scanning the calibrator QR code;

Quality Control (optional): 1 bottle each of high and low points, containing BSA buffer with different amounts of LH antigen added. The target range of the quality control is obtained by scanning the quality control QR code.

Note ①: Reagents from different batches of kits are prohibited from being used interchangeably.

Note ②: Six calibrator points or two calibrator points are optional. If calibrators and quality controls have been provided to the user, the kit does not need to be reconfigured.

2. Companion Reagents

The following companion reagents are not included in this product but are required for detection:

Pre-excitation solution and excitation solution: Excite acridinium ester to catalyze the production of photons, so that the chemiluminescence intensity can be detected using a chemiluminescence immunoassay instrument.

Washing solution: Diluted for washing the reaction system.

Source: Provided by our company.

Note: To ensure the accuracy of the test results, the manufacturer of the companion reagents should not be changed arbitrarily during the test.

3. Calibrator Traceability

The calibrator is traceable to the "National Standard Substance for Luteinizing Hormone (LH) Immunoassay" provided by the China National Institute for Food and Drug Control.

【Storage Conditions and Shelf Life】

The reagents are stable for 15 months at 2-8℃, protected from light, and should not be frozen.

After opening, liquid reagents are stable for 30 days at 2-8℃, protected from light.

【Production Date】

See product label.

【Applicable Instruments】

Cosmed SMART500S, 6500, or Tuochuang Medical TC-300 fully automated chemiluminescence immunoassay instrument

【Sample Requirements】

1. This kit is suitable for serum samples.

2. After blood collection, serum should be separated immediately for analysis to avoid hemolysis. Testing must be completed within 24 hours at room temperature, stable for 3 days at 2-8℃, and stable for 30 days at -20℃. Avoid repeated freezing and thawing.

3. Frozen and refrigerated samples should be returned to room temperature and mixed thoroughly before use.

4. Avoid using samples with severe hemolysis or high concentrations of lipemia.

5. The sample volume should be no less than 200μL.

【Test Method】

Note: Please read the instructions carefully before use. Working conditions: Room temperature (15～35℃); Relative humidity ≤80%.

1. Pre-test preparation

1.1 Before the test, all reagents should be placed at room temperature, and the instrument should be preheated for at least 30 minutes;

1.2 According to the system operating instructions, perform the loading of reaction tubes (cups), the addition or replacement of matching reagents, waste liquid, waste tube cleaning, and liquid path filling operations.

2. Reagent and sample loading

2.1 Before reagent loading, place the reagents to be used on the mixing equipment for thorough mixing. Visually inspect the reagent solution components; they should be clear, free of foreign matter, precipitates, and flocculent matter. The magnetic microparticle reagent should be a uniform suspension without obvious aggregation;

2.2 According to the system operating instructions, scan the reagent barcodes to complete the loading of magnetic microparticle reagents and anti-reagents;

2.3 After mixing the calibrators and quality control samples, transfer them to the instrument's dedicated reaction cups and load them into the instrument's sample positions. After scanning, load the centrifuged serum samples directly into the instrument's sample positions.

3. Detection steps

For optimal detection performance, please follow the relevant instructions in this manual and operate according to the operating manual of the fully automated chemiluminescence analyzer.

Note: The detection reaction process and related parameters have been predefined in the instrument operating software.

4. Calibration curve and calibration

The calibration curve is obtained by scanning the main curve card to generate it directly, or by directly detecting six calibrator points to generate a fit, and the system also supports obtaining a working calibration curve using a two-point calibration method.

Note ①: Due to inter-instrument differences and system differences caused by different operators, operating environments, and matching general reagents, the six-point direct calibration method is recommended to obtain the working calibration curve.

Note ②: Due to reagent activity drift and changes in batches of general reagents, the calibrated working curve needs to be recalibrated after a certain period of use. Recalibration should be performed when the following situations occur:

l After 1 month (28 days) of using the same batch number of reagents;

l When using a new batch number of reagents, the whole box or luminescence pre-excitation liquid and excitation liquid;

l When the quality control values are outside the quality control range;

l After each maintenance of the analyzer.

5. Result output

The analyzer automatically calculates the LH concentration of each sample using a working curve obtained by two-point calibration from the standard curve, and the results are expressed in mIU/mL.

6. Quality control

6.1 Two levels of quality control samples should be determined daily along with sample testing, treating the quality control samples as patient samples;

Note: It is recommended to use at least two levels of commercial quality control products. If the test results are within the system's acceptable quality control range or within your specified range (determined by an appropriate internal laboratory quality control program), the test results are satisfactory; otherwise, it indicates that the test results are unreliable and a test report should not be issued.

【Reference Range】

Note: The LH levels measured will vary depending on the region, individual, and method used for testing. Therefore, we recommend that each laboratory establish its own normal value range. A diagnosis should not be made based solely on the LH value obtained by this method; the results should be analyzed in conjunction with other clinical data, including the patient's specific condition and treatment status.

【Interpretation of Test Results】

1. Due to methodological or antibody specificity reasons, testing the same sample using reagents from different manufacturers may yield different results. Therefore, results obtained using different reagents should not be directly compared to avoid erroneous medical interpretations.

2. Test results exceeding the test range of the kit are calculated results obtained by extrapolating the calibrator curve. When reporting such results, please pay special attention. If an accurate measurement is desired, dilute the sample appropriately before testing; the maximum dilution factor is 100 times.

3. Quality control products can serve as a reference for the reliability of the results of the current experiment; their measured values should be within the allowable range specified on the quality control sheet for this batch of products. Test results should be judged comprehensively based on the reference range and other clinical factors and results. When the test result is close to the upper limit of the reference range, consider performing a confirmatory test on the sample.

【Limitations of the Test Method】

1. Severe hemolysis (hemoglobin ≥500mg/dL), lipemia (triglycerides ≥1500mg/dL), jaundice (bilirubin ≥20mg/dL), and contaminated samples may affect the test results and should be avoided;

2. The test results of this kit are for clinical reference only and cannot be used alone as the basis for confirming or excluding cases. To achieve diagnostic purposes, this test result should be used in conjunction with clinical examinations, medical history, and other examinations.

3. This product can be used for the determination of human serum samples, and the reliability of determining LH concentration in other body fluid samples has not been fully confirmed.

【Product Performance Indicators】

1. Reagent Performance Indicators

1.1 Accuracy: The ratio of the measured value to the labeled value of the calibrators in the kit should be between 0.900 and 1.100.

1.2 Precision: Intra-batch precision ≤8%, inter-batch precision ≤10%.

1.3 Blank limit: Should not exceed 1mIU/mL.

1.4 Linearity: In the range of (1～200) mIU/mL, the correlation coefficient r should be ≥0.9900.

1.5 Specificity: The measured result of human chorionic gonadotropin (Total-hCGβ) with a concentration not less than 1000IU/L should not be higher than 1mIU/mL; the measured result of thyroid stimulating hormone (TSH) with a concentration not less than 200mIU/L should not be higher than 1mIU/mL; the measured result of follicle stimulating hormone (FSH) with a concentration not less than 200IU/L should not be higher than 1mIU/mL.

2. Calibrator Performance Indicators

2.2 Accuracy: The ratio of the measured value to the labeled value of the calibrators in the kit should be between 0.900 and 1.100.

2.3 Uniformity: Inter-bottle difference (CV) ≤8%.

3. Quality Control Product Performance Indicators

3.1 Expected results: The test value should be within ±15% of the target value of the quality control product.

3.2 Uniformity: Inter-bottle difference (CV) ≤8%.

【Precautions】

1. This reagent is for in vitro diagnostic use only. Reagents from different batches are prohibited from being used interchangeably. Do not use expired reagents.

2. Some components of the kit contain bovine-derived materials, but these raw materials are not from countries or regions with bovine spongiform encephalopathy (BSE) epidemics, and their production is not carried out in countries or regions with BSE epidemics. If it is bovine serum, it will be filtered and sterilized after purchase before being used in kit production. Therefore, the kit does not pose a risk of transmitting BSE; some components of the kit contain porcine-derived materials, but these raw materials are not from countries or regions with foot-and-mouth disease epidemics, and their production is not carried out in countries or regions with foot-and-mouth disease epidemics. If it is porcine serum, it will be filtered and sterilized after purchase before being used in kit production. Therefore, the kit does not pose a risk of transmitting foot-and-mouth disease; this reagent does not contain human serum, but should still be considered a potential biohazard.

3. This reagent contains preservatives. Avoid contact with skin. If skin accidentally comes into contact with the reagent, rinse thoroughly with plenty of water.

4. All reagents should be stored at 2～8℃, avoid freezing at -20℃, and equilibrate to room temperature before use.

5. The magnetic microparticle reagent should be thoroughly mixed before use, and strong shaking should be avoided.

【Explanation of Labels】

Store at 2-8℃ Avoid direct sunlight Store upright

In vitro diagnostic reagent Production date Expiration date

  Product batch number See instructions for details

【References】

1. Shivapathasundram, Ganeshwaran, Kwik, Michele, Chapman, Michael. Luteal phase defect: part of the infertility zeitgeist or relic from the past?. Human Fertility, 14(1):60-63.

2. Shang Wei, Wang Aiming, Shu Mingming, et al. The relationship between luteinizing hormone level and recurrent miscarriage. Hebei Medical University Journal, 2012, 33(11):1275-1275.

3. Chen Yanru, Lin Junmin, Hou Quanling, et al. The application of luteinizing hormone in ovulation induction. China Maternal and Child Health Care, 2018, 033(004):951-954.

【Basic Information】

Registrant/Manufacturer: Zhejiang Tuochuang Medical Technology Co., Ltd

Address: No. 1 Building, Thousand Talents Plan Industrial Park, No. 78, Zhancheng Avenue, Zhuji City, Zhejiang Province

Production address: No. 1 Building, Thousand Talents Plan Industrial Park, No. 78, Zhancheng Avenue, Zhuji City, Zhejiang Province

After-sales service unit: Zhejiang Tuochuang Medical Technology Co., Ltd

Production license number:

Tel: 0575-87178692

Fax:

Post code: 311800

Website: www.zjtcyl.com

【Medical Device Registration Certificate Number/Product Technical Requirements Number】

【Approval Date and Revision Date of Instructions for Use】

Approval date:

Revision date: