MTHFR C677T GENETIC TESTING KIT
The product contains five invention patents with world-leading core technology and significant clinical application value. The kit has been recognized as a national innovation medical device and can be operated by an average testing personnel in hospital.
1. Product Features:
● Fast: The genotype of the kit can be read in 5 minutes after PCR.
● Convenient: GoldMag Lateral Flow Assay technology allows one-step reaction for simple operation for 50 μL PCR product.
● Accurate: Over 99% general coincidence rate achieved with the sequencing method.
● Universal: Testing can be conducted in all levels of health and medical institutions with no special instrument and equipment needed.
2. Product Usage:
The product, designed for MTHFR C677T genotype testing, can provide auxiliary diagnosis for high homo-cysteine levels and folate metabolism disorder patients. It can also provide clinical reference for giving folic acid supplement. The testing of MTHFR can be used to screen varieties of critical diseases so as to implement interference in advance for disease occurrence reduction. The product is suitable for the physical examinations of the patients of cerebrovascular disease, pregnant women, middle-aged & elderly population, and smokers or normal people.
● Providing complete market access procedures and certification services for authorized dealers.
● Providing innovative and competitive core products to dealers /agents.
● Continued new product launching to meet the changes and demands of the market.
● Providing comprehensive training and technical support services to assist dealers in market development.
● Being in compliance with laws and regulations; honest and trustworthy; having a strong sense of cooperation; capable of keeping commercial secrets.
● Being a legal person as an enterprise or a natural person with full civil capacity; in compliance with market management systems and implementation of market policy.
● Having strong cognitive and learning capability; with great interest in protein biomarkers and genotyping testing industry.
● Owning mature network resources and sales team in the regional market; or having extensive market operation experience with strong market development capabilities.
The problems existing in the process of using MTHFR C677T Genetic testing kit
Q1：What are the requirements for clinical samples when using MTHFR C677TGenetic testing kit? A1:Genomic DNA isolated from fresh whole blood should be obtained before using this series of kits. Protocols of venous blood and genomic DNA isolation are shown below:(1)200ul—1ml Venous blood(Containing EDTA anticoagulant ) If it was stored at 2-8 °C, and extracted in 7 days;if more than 7 days it had better be frozen; the freeze-thaw cycles cannotbe processed more than 5 times.(2) DNA concentration should be greater than 5ng/ul; A260 / A280 should be between 1.6-2.0(3) If the sample contains a clot, itrequired in extracting DNA after homogenization or grinding .
Q2：Which result should be considered as nullity? How to deal with? A2:(1) When the negative control experiments show strip in the test line,the results cannot be trusted.
It may be caused by improper operation or contamination, so we need to exclude contamination and re-operate.
(2) When the positive control experiments show no strip in the test line,the results cannot be trusted.
It may be caused by improper operation or problems reagents
, so we need to accord to the instructions and re-operate.
(3) When the control line show not strip,the results cannot be trusted. It may be caused by improper operation or the deterioratedKit components. You can replace the text card and repeat the experiment again.
Q3: How to effectively prevent contamination in the process of PCR? A3:(1)Laboratories should follow the specifications ,namely partition PCR laboratory operations;PCR laboratory partitioning;
Items are special districts;Pipette plus templates and primers can not be mixed;To be replaced after each sample tips.
(2) Before the experiment, please briefly centrifuge
(3) Use disposable gloves during the experiment and timely replacement
(4) Before and after the experiment, using ultraviolet light test rig 30min
Q4: How to store the kit to make it in the best condition? A4:Test card, positive control card and negative control card had best be stored at Dark and dry place, ban frozen.
After unpacking, test card will fail due to damp, so please use within one hour.
If the environment is higher than 30 degrees or a high humidity, you had better immediately use after open it
PCR reaction kit save more than 3 degrees, no more than five times repeated freezing and thawing
PCR reaction kit save at -20℃ , freezing and thawing no more than repeated five times. Validation for 12 months.
After opening the components of PCR reaction kit ，save no more than a week at 2-8℃ and -20℃ can keep to the validity
Q5：Are there genetic test results timeliness? Whether different times detect the results is differ? A5:Human single nucleotide is a marker, soit is a very small changing probability in the life
Therefore, the results of genetic testing has not timeliness .if you once detected, it will be valid for all of your life. It will not change in different periods
Q6: How the text of MTHFR C677T pointdetection guide to use
folic acid? A6:Methylenetetrahydrofolatereductase(MTHFR) CT mutant gene type
Gold magnetic immunodiagnostic kits FAQ
Q7: What circumstances is invalid interpretation of the results? How to deal with? A7:Qualitative products:
There are a lot of randomness when the judgment result, the judgment time is not uniform because staff in the use of products distributed in different departments and are not ehe same level,especially resulted from the different working environment, temperature, humidity. People always ignored the effective time developed by manufacturers when randomly judging results according to the time stamp line color . Therefore, people need to strictly follow the provided instructions.when testing .
Control line (C line) does not appear bands, the results can not be trusted,because the operation may be incorrect or the strip were damaged, replace the strip and retest. If the problem is not resolved, please contact the supplier or manufacturer.
No judgeing mistakes when using quantitative products due to the instrument will automatically calculate the interpretation of measurement resultswithin the stipulated time.But when the control line (C line) does not appear bands, the results can not be trusted, resulted from not operate correctly or strip is damaged, replace the strip retested. If the problem is not resolved, please contact the supplier or manufacturer.
Q8: What are the requirements for test sample? A8:Serum samples:drawn venous blood , add the whole blood sample to the sample collection tubes containing no anticoagulant, 37 ℃ water bath for 20-30 minutes (or set for at least 30 minutes at room temperature), after fibrinogen be fully coagulated ,then centrifugate (1500-3000rpm / min), detected after separation of serum within 4 hours.
( available according to kit instructions request) extracted from venous whole blood, anticoagulant (EDTA, citrate and heparin, can be used as an anticoagulant) is added to the whole blood sample in sample collection tube, the plasma is detected within 4 hours after centrifugated.
If fresh serum or plasma can not be detected immeditately, it should be stored at 2-8 ℃, deposit period of not more than three days, or placed in -20 ℃ cryopreservation, valid for six months, not to repeated freezing and thawing.
Q9:what impaction will appear if the kit save improper? A9:When the kit did not follow the prescribed storage conditions to save, it cause inaccurate test results or test strip does not appear, thus it should be strictly in accordance with the requirements of saving use. Save it in the following conditions: 4-30 ℃ stored in a cool dark place, no frozen; valid for 18 months. After unpacking detection reagent ,it will be valid due to moisture,thus use it within one hour; people should be ready to use as soon as possible where the temperature is above 30 ℃ or in high humidity environments.
The production of isolated genomic DNA Frequently Asked Questions
Q10:The Isolated DNA amount is too small or the purified DNA purity is too low A10:The possible causes and recommendations
(1)Before DNA purification, Confirming cleaning fluid BW-I and cleaning fluid BW- II have been correctly formulated according to the instructions .
(2)confirming the magnetic particles has been adequately shake before using. At the same time, please do not freezing dry andCentrifugal the magnetic particles
(3)Thetime of blood storageis too long.Do notsavemore than a week at 2-8℃. When you need long-term storageplease store at-80 ℃.
(4)Do not place a long room temperature with proteinase K and avoid repeated freezing and thawing. If you need to multiple use,
please dispense the reagent .
Q11: Isolated and purified DNA concentration is too low A11:Shocking to the magnetic particles can be fully dispersed state while reducing the elution volume.
GMag-Sep 8 automatic accounting extraction instrument and supporting RNA extraction kit for use of common problems
Q12: Isolation and purification of DNA small amountandlow purity A12:The case caused There are many reasons
1 Blood preservation may be a long time, 2~8 degrees do not save more than a week, if the long-term preservation of advice is placed in the -80 degrees Celsius cryopreservation
2 Reagent strips of storage, in accordance with the requirements of the storage needs, otherwise the temperature is too high or too low magnetic particles and reagents might cause a lot of damage.
3 Protease K to save, do not place a long time and to avoid repeated freezing and thawing at room temperature.so as not to affect its activity. Such as the need for repeated use, can according to the experimental parameter will be packing
Q13: reagent strip or T-shelf rack with liquid A13:A O-ring wear and leaks occur, you need to replace the O-ring
When Parallel test hole with a gun head absorb liquid, liquid surface height is not a shot in the head
A description of the gun head suck the liquid volume does not meet the requirements, the reason may be;
1 pipette tip deformation needs to be replaced
2 O-ring damage, referring to the suction device to check for tips to learn the level of the solution to be checked
1. Putative mechanisms for vascular damage by homocysteine.
作者：Bellamy M F, McDowell I F.
出处: Journal of Inherited Metabolic Disease. 1997, 20(2): 307-15;
注: ((Journal of Inherited Metabolic Disease》杂志主要致力于先天代谢异常的研究，具有国际性和多科学性。该杂志的文章、论文、病理报告涉及人类和高等动物代谢异常方面。
2. Homocysteine induces congenital defects of the heart and neural tube: effect of folic acid.
作者: Rosenquist TH, Ratashak SA, Selhub J.
出处: Proc Natl Acad Sci USA. 1996, 93(26): 15227-32;
注: (Proceedings of the National Academy of Sciences of the United States of America)是美国科学院的院刊，于1914年创刊，提供具有高水平的前沿研究报告、学术评论、学科回顾及前瞻研究性论文以及美国国家科学学会学术动态。PNAS收录的文献涵盖生物、物理和社会科学。
3. Acommon mutation in the 5,10-methylenetetrahydrofolate reductase geneaffects genomic DNA methylation through an interaction wish folate status
内容摘录：甲基化是DNA分子重要的表观遗传特征，MTHFR催化5一甲基四氢叶酸的合成,促进同型半胱氨酸生成甲硫氨酸，为后续反应提供甲基。本研究根据MTHFR基因型选取105名突变纯合子个体 (TT)和184名野生纯合子个体(CC)为研究对象，采用液体色谱/质谱法评估外血基因组DNA甲基化水平。研究发现，基因组DNA甲基化水平与叶酸浓度直接相关，与血浆总同型半胱氨酸(tHcy)的水平((P<0.01)成反比:相比CC野生型个体，TT基因型患者DNA甲些化水平降低，TT个体的DNA甲基化与红细胞叶酸的甲酰化呈负相关。结果表明，MTHFR C677T基因基突变通过与叶酸的相互作用影响DNA的甲基化。
作者:Friso S, Choi SW, Girelli D, et al.
出处:Proc Nat! Acad Sci USA. 2002, 99(8): 5606-11.
4. A common mutation in the methyl enetetrahydrofolate reductase gene is asso-
作者: Bagley P J, Selhub J.
出处: Proc Natl Acad Sci USA. 1998, 95(22):13217-20.
5. MTHFR 677C→T genotype is associated with folate and homocysteine concentra-tions in a large, population-based, double-blind trial of folicacid supplement.
内容摘录: 本文研究了MTHFR C677T基因突变对中国人叶酸转化能力的影响，研究得出以下结论: 叶酸补充后，至少需要3个月才能达到一个稳定的血药峰值，因此需要孕前补充叶酸;
作者：Crider K S, Zhu J H, Hao L, et al.
出处:The American journal of clinical nutrition, 2011，93(6): 1365-72;
注:(The American Journal of Clinical Nutrition》是评价最高的同行评议杂志，其文章包括世界上关于人类营养的基础和临床最新的研究文献，这些研究涉及肥胖、维生素和矿物质、营养和疾病以及能量代谢。AJCN被专业图书馆协会((SLA)选为过去一百年里生物学和医学领域最具影响的100种期刊之一。
6. Relationship between genetic polymorphisms of methyl enetetrahydrofolate reduc-tase (C677T, A1298C, and G1793A) as risk factors for idiopathic male infertility.
内容摘录: 确定了MTHFR基因3个位点的突变(C677T, A1298C和G1793A)、血清叶酸、血浆总同型半胱氨酸(tHcy)水平与男性的生育能力状况包括精液参数的关系。本研究采用聚合酶链反应一限制性片段长度多态性(PCR-RLFP)技术对164名原发性不育男性和328名健康男性确定MTHFR的基因型，并测定了血清总同型半胱氨酸水平、叶酸和维生素日B12的浓度。结果显示，MTHFR677T等位基因携带者大大增加了患男性不育症的风险，同时携带677T、1298C,1793G等位基因的男性表现出高半胱氨酸和低叶酸水平的特征。
作者: Safarinejad MR，Shafiei N，Safarinejad S.
出处：Reproductive Sciences. 2011，18(3): 304一15.
7. Abnormal folate metabolism and mutation in the methylenetetrahydrofolate
内容摘录: 携带M丁日FR C677下突变的母亲比未携带该突变的母亲生出唐氏患儿的概率高2.6倍(95% CI:1 .2，5.8; P<0.03).
作者: James S J，Pogribna M，Pogribny I P, et al.
影响因子: 6 .50;
出处: The American journal of clinical nutrition. 1999, 70(4): 495一501.
8. Does the interaction between maternal folate intake and the methylenetetrahy-drofo-late reductase polymorphisms affect the risk of cleft lip with or without cleft palate?
作者:van Rooij I A, Vermeij-Keers C, Kluijtmans L A, et al.
出处:American journal of epidemiology. 2003, 157(7): 583-591.
注:((American Journal of Epidemiology))杂志是流行病学领域领先的杂志，涵盖流行病学研究和方法.旨在为专业人士和公共卫生工作者提供专业文献。
9. Risk of congenital heart defects is influenced by genetic variation in folate metabolism.
作者:Christensen KE, Zada YF, Rohlicek CV, et al.
出处:Cardiol Young. 2013, 23(1):89-98.
注:(Cardiology in the Young)致力于影响儿童的心血管问题，主要集中于先天‘!生心脏病领域，目前杂志已涉及了该领域的所有方面。杂志内容涉及研究的许多形式，本杂志最大的特色莫过于其文章图表的可视化。
10. Epistasis between COMT and MTHFR in maternal-fetal dyads increases risk for preeclampsia.
内容摘录: 先兆子痫是围产儿发病率和死亡率的首要原因。这种疾病除了与多个基因有关外，还受环境和社会因素等因素影响。研究结果表明先兆子痈和孕产妇和胎儿的COMT与MTH FR C677T基因突变有极高的相关性。
作者: Lori D. Hill, Timothy P. York, Juan P. Kusanovic, et al.
出处: PLOS One. 2011，6(1):e16681.