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This section is a non-exhaustive guidance on technical encountered questions on PCR and FISH related to our products. If you are experiencing issues with your FISH probes or protocol the FAQ section may feature the solution to your problem. For any question not mentioned below, please do not hesitate to contact us.

 

Is there any conflict between CYP2C19 gene test and the existing clinical platelet function test?

There is no conflict. Clopidogrel gene testing can be used to predict the enzyme activity encoded by the CYP2C19 gene before the patient's medication, and according to the enzyme activity of the subject adjust the dosage and regimen. For high-risk patients key monitoring should be led before any medication or medication change. At the present stage, the main reasons affecting the drug treatment effect include genetic factors, cellular factors (such as platelet renewal and acceleration may reduce the therapeutic response to antiplatelet agents), clinical factors (dosage, liver and kidney dysfunction, drug interactions), and other clinical platelet function tests are cytological factors that affect the drug’s efficacy. Clinical studies have shown, after excluding cellular and clinical factors, genetic factors are the main cause of adverse reactions, and are independent risk factors. So the genetic testing is not in conflict with the existing clinical testing.

 

What are CYP2C19 gene polymorphism detection advantages in drug selection?

- To enable patients who are treated with Clopidogrel to understand the risks of their own treatment before treatment and to avoid serious adverse reactions as early as possible in patients ' own body drug trials.

- CYP2C19 gene testing, a single test that can benefit a lifetime and can help patients who do not carry a functional allele deletion to save unnecessary expenses.

- Patients with weak metabolic ability or not eligible for Clopidogrel clinical treatment can be adjusted as early as possible to avoid the occurrence of adverse clinical events such as stent thrombosis.

- To provide the necessary reference for doctors to select other medication for patients.

 

What is the duplicates detection rate of our kit?

Duplicates detection rate refers to single test success rate. For some products, at the final detection stage, there will appear a reading signal failure situation leading to a new test process, increasing labor and reagent costs.

Till now, our products have not received any complain from any customer related to results detection. Theoretically, as long as the sample DNA concentration and purity are not of a concern, there will not be genotype detection issue. HealthCare Biotech product is sure.

 

How to explain cost of genetic testing to allow patients to gain net benefit from it?

The medicine can be used regardless of social status. It has high efficacy and low side effects. New drugs are usually more expensive, with less available research data on efficacy and toxicity, and the impacting factors are not clear, which need to be studied and confirmed in long-term clinical use. In contrary, old medicine has been used for a long time, with a large number of clinical use, and confirmed safety and effectiveness.

 

How many years can gene testing be done?

Generally, it is a lifelong benefit, under normal conditions human genes cannot mutate.

 

How to get test results from the LIMS system of the hospital or the laboratory? Or is there any software that can report the test directly?

Currently, there is no existing software. We can provide the genotype test report (Word version) and technical support to the customer/user to help to check the test results on the system.

 

The DNA added in sample amount is too large, and the DNA amount is not enough

It is recommended to add 10μl to each hole.

 

Does the substitution by other products can replace Statin detection problem?

Our Clopidogrel and Warfarin kits can run with other PCR programs, but using HealthCare Biotech Statin will be the best option.

 

How long does the blood sample be able to do PCR detection?

Fresh blood samples, EDTA anticoagulant frozen preserved within a month.

 

What is FISH?

Literally Fluorescence In Situ Hybridization is a technique used in pathology based on gene testing to guide target therapy and tumor typing.

 

How does the FISH process work?

The process consists of paraffin sections dewaxing, permeating, enzymatic digestion, probes hybridization, staining (dyeing), and examination by fluorescence microscopy.

 

What equipment does FISH need?

Fluorescence microscope and hybridization instrument.

 

What the pretreatment equipment do?

Introduce FISH operation, perform pretreatment before hybridization

 

The curve has no start line, but CT value. Can it be removed through parameter settings?

Generally cannot be eliminated through parameter settings, but only manually by setting an elimination threshold line.

 

What does 681G>A mean?

The CYP2C19 gene at 681 locus mutates from one ribose nucleotide to another, and specifically manifested by Guanine mutation into Adenine.

 

Compared to similar products, what are HealthCare Biotech product advantages?

Methodological advantages: closed reaction, reduced operational steps, simple and easy to use, no special equipment required and easily found in hospitals and laboratories;           

Product advantages: eighth tube packing, seal up paraffin, good reagent stability.

 

HealthCare Biotech product characteristics?

Short probes, no repetitive sequences, high hybridization efficiency, 2h hybridization.

 

The CT value is too high

First, look at the positive CT value, if the positive control CT value is normal, then the sample concentration is too low, or the sample purity is too low. If the positive CT value is also high, it may be related to a reduced PCR enzyme activity.

 

The expansion magnitude is low

First, check the positive control whether is normal, if the positive control is normal, the sample purity is low. If the positive control is also low, it may be the instrument channel problem, check and fix the instrument.

 

There is no amplification curve

First, check whether there is amplification of the positive control, if there is amplification, it is a sample problem. If there is also no positive control amplification curve, the issue may be related to PCR enzyme inactivation.

 

The positive control FAM channel amplification curve amplification intensity is low?

Check if all samples, including positive control, have low FAM amplification intensity, relevant to instrument channel failure, check and fix the instrument.

 

Positive control amplification?

If one of the two samples is amplified, it is contaminated by the experimental operation or aerosol pollution; if all the negative controls are amplified, the negative control reagent is contaminated.

 

How to deal with not enough DNA quantity?

Can dilute DNA and DNA concentration should not be less than 1ng/μl.

 

How the product is transported?

Cold chain logistics transportation.

 

Nonspecific amplification?

Too much DNA addition, heterologous sources of DNA contamination.

 

What equipment does FISH experiment need?

Hybridization apparatus, water bath pot, fluorescence microscope

 

What does FISH do?

Adjuvant diagnosis, tumor prognosis, target medication, etc.

 

No dispersed cells are observed on tissue under microscope

Inadequate pretreatment of digestion enzymes.

 

How to deal with strong fluorescence background signal?

The slides are not clean and the hybridization washing is insufficient.

 

Weak hybridization fluorescence point signal?

Improper pretreatment, inadequate denaturation during hybridization and improper hybridization conditions

 

Fast Probe and why fast?

Because of the absence of non-repetitive sequences.

 

Is there any fully FISH automatic processing system (Slides heating ----- Hybridization ----- Washing)?

Not at the moment, but this fully experimental automated processing system is the next generation of our R&D objective.

 

How HealthCare Biotech Fish experiment figures/pictures effect is compared with other manufacturers?

Compared with some brands such LBP and Abbott, our fast-probe hybridization time is short to 2 hours with clear hybridization fluorescent spots, and the overall effect was the same as that of their overnight hybridization.

 

Does the FISH diagram go through the P-graph of software P?

Our FISH diagram does not go through the P-graph of software P since its contrast on the computer is very clear.

 

Do HealthCare Biotech pretreatment machine will be affected if the slides thickness was different?

In general, for the same batch we process, there will not be any big difference on the slides thickness, and we do not put different slides from different batches to constitute a batch. In addition, our machine according to the user thickness setting can determine the corresponding enzymatic digestion time.

 

What are the requirements for blank slides when using HealthCare Biotech’s pretreatment system?

Puncture or embedded cells/tissues can be used, the slides thickness generally has no particular requirements, and the normal situation quite is good.

 

Can HealthCare Biotech reagent be used only in conjunction with the apparatus? Or does the apparatus need to be matched with the reagents?

Reagents being used either manually or in conjunction with our instrument, the entire step time remains similar. Our instrument must be used with our reagents, because being programmed according to our own reagents, and if different reagents are used, we cannot guarantee the effect of the pretreatment.

 

Can reagent be tested for comparison?

Samples can be provided free of charge and the customer takes on the freight fee.

 

What is the reagent consumption of each slice at the pretreatment and the corresponding price?

The average reagent consumption during pretreatment is about 13ml/slide, for a related price around $1.50.

 

How to process if after checking slides digestion stage during pretreatment one would like to lengthen the digestion time?

In the case the enzyme digestion needs to be checked, the machine can be set to the semi-automatic mode. The instrument will automatically remind by an alarm sound to set the digestion time and digestion stage can be observed as well as digestion can be interrupted by pressing on “Stop” button.