Application Of Scientz Tissue Grinder In The Processing Of Afric An Swine Samples

Pig Eight Rings: Monkey Brother save me!

Wukong: nerd, what is flustered!

Pig Eight Rings: Monkey Brother, you haven't heard of it! African pigs are coming, one is infected, and one nest is suffering! I have heard that many brothers in Liaoning, Heilongjiang, Jilin, Jiangxi, Fujian, and Sichuan have been culled.

Wukong: nerd! That is not far from me!

Question 1. What is African swine fever?

      African Swine fever (ASF) is an acute, hemorrhagic, and severe infectious disease caused by African Swine fever virus (ASFV) infection of domestic pigs and various wild boars (non-infected). It is characterized by a short onset, with a mortality rate of up to 100% for the most acute and acute infections, and can only be confirmed by laboratory monitoring.

       On August 3, 2018, China confirmed the first case of African swine fever.

Question 2: Introduction to sample processing methods for African swine fever detection

I. African piglet sample processing

1. Purpose

Standardize the processing procedures for ASFV samples and experimental materials to ensure the sequential development of ASFV-related experiments and biosafety in laboratories.

2. Scope of application

Pre-experimental treatment for suspected ASFV clinical samples

3. Procedure

3.1 Experimental materials

ASF susceptible animals, mainly domestic pigs, wild boars and crickets, usually collected samples of domestic pigs, wild boar blood samples, tissue samples and cockroaches. Samples collected in the field usually require simple pre-treatment for subsequent testing and diagnostic work.

Serum: for viral nucleic acid detection, viral antigen and antibody detection. The syringe or blood collection tube of the collected pig blood is tilted at room temperature for 2 to 4 hours (preventing exposure), or placed in a 37 ° C incubator for 1 h. After most of the serum is precipitated, the serum is taken out, and serum is separated by centrifugation if necessary. . (Minimum recommended amount 5ml).

Anticoagulated whole blood: used for viral nucleic acid detection and viral antigen detection.

The anticoagulant is added proportionally in a vacuum blood collection tube or syringe prior to blood collection. Collect blood and gently invert it upside down to mix the blood and anticoagulant to prevent blood from clotting while avoiding hemolysis. It is also possible to put the blood into a sterilized bottle containing glass beads and shake off the fibrin. If necessary, a double antibody can be added to inhibit bacterial contamination that may occur during blood collection or blood collection.

When blood samples are used for viral nucleic acid detection, EDTA is generally used as an anticoagulant without heparin. (Minimum recommended amount 1mL).

Tissue samples: in vivo collection, using tonsils, for clinical health animal ASF monitoring.

Autopsy ASF infected animals or dead animals were dissected, including: ASFV target organs collection: spleen, lymph nodes, kidneys, tonsils.

Collection of other pathological organs: Collect organs such as liver, lung, heart, etc. with obvious lesions, and select the site at the junction of disease and healthy tissue.

Sample collection of corrupted animal carcasses: The femur was taken by necropsy, and the attached muscles and ligaments were all removed, and bone marrow was collected. The minimum recommended amount is 5 grams.

The collection of cockroaches: cockroaches can carry out nucleic acid detection, antigen detection and morphological identification of cockroaches, find the wall edge, wall seam, wooden or tile roof of the pig pen, or find the cockroach on the ground of the pig pen, or view the pig Epidermis, collecting sputum.

Note: Samples used for virus isolation and pathology testing are prohibited from storage at -20 °C.

3.2 Laboratory equipment and reagents

1) Desktop high-speed refrigerated centrifuge, tissue grinder, water bath, timer, refrigerator and pipette.

2) Sterile scissors, forceps, steel balls, 2 mL EP tubes, pipette tips (10 μL, 200 μL, 1000 μL), etc.

3) Other reagents and consumables mainly include: 0.01mol/L PBS (pH 7.2), 0.8% NaOH, 75% alcohol cotton ball, and the like.

3.3 Sample Processing Procedure

3.3.1 Serum and whole blood sample processing

Take 1 mL of serum or whole blood sample in a sterile EP tube and save for use.

3.3.2 Tissue sample processing

It is suitable for the treatment of tissue samples such as spleen and sputum. Reduce the pollution as much as possible. For each tissue block or sputum, use a sampling device such as a flame disinfection scissors. The tissue blocks should be placed in a sterilized container and sealed immediately, and marked to prevent mutual contamination between tissues.

3.3.2.1 The handling of tissue samples should be carried out in a secondary biosafety cabinet. Do not allow liquid to spill into the biosafety cabinet.

3.3.2.2 Put 0.1-0.2g tissue block into 2mL EP tube, cut into pieces with scissors, add 1-2mL 0.01mol/L PBS (pH 7.2), then add 1-2 sterilized steel balls, after sealing The surface is disinfected for use. The remaining sample is returned to the container, resealed, and the surface is sterilized and stored in a -70 ° C refrigerator.

3.3.2.3 Fragmentation of tissue samples The tissue can be manipulated outside the biosafety cabinet to make the tissue into a tissue suspension.

Heavyweight recommended Scientz-48L frozen high-throughput tissue grinder

3.4 Virus inactivation

The inactivation of the virus should be carried out in a secondary biological safety cabinet. The treated serum, whole blood or ground and broken tissue samples were placed in a 60 ° C water bath together with an EP tube and allowed to stand for 30 min to inactivate.

3.5 Instrument processing after use

Scissors, tweezers, etc. should be placed in a sterilization tank for disinfection, placed in an iron lunch box, and placed in a sealed bag for autoclaving. The container containing the tissue sample preservation solution and the centrifuge tube containing the tissue block should be sealed, and the surface is sterilized and placed in a sealed bag for autoclaving. The steel balls, tips and other wastes are immersed in 0.8% NaOH for 30 minutes, and then placed in a sealed bag for autoclaving. The disinfectant residue should be autoclaved in a sealed container.

3.6 Precautions

Before operation, confirm that the biosafety cabinet and laboratory have been thoroughly sanitized to prevent cross-contamination between samples.

Second, the nucleic acid detection of African swine fever virus can be carried out by real-time fluorescent PCR, and the specific operation steps are subject to the guide of the instrument production company.

Question 4, virus killing and prevention and control methods

Although the African swine fever virus is relatively stable in the environment, it is easier to inactivate during the cooking process of pork. When heated at 70 °C-75 °C for more than 30 minutes, the virus will be killed, which means that we only need to cook during daily cooking. Cook the pork thoroughly (including temperatures in the middle of the part at least 70 ° C), even if there is a virus will quickly lose the ability to infect and lose activity.

Introduction of Scientz Bio Scientz-48L Frozen High-throughput Tissue Grinder

The Scientz-48L frozen high-throughput tissue grinder from Shinzo Bio has the following features:

1. Various application scenarios. Different tissues can be dry-grinded, wet-milled, mixed, and homogenized.

2. The experiment is highly efficient. It can improve the efficiency and quality of nucleic acids in large-flux samples.

3. The experimental process is safe. The grinding process uses a closed disposable centrifuge tube, which effectively avoids cross-contamination of the sample.

4. The instrument has good stability. The same grinding procedure can be set for the same tissue sample to achieve the same grinding efficiency and improve the repeatability of the experiment.

5. The grinding effect is good. The product is ground with high-efficiency grinding balls, which are more uniform and sufficient for grinding the sample compared to the traditional grinding method.

At present, scientz Biotechnology Scientz-48L frozen high-flux tissue grinder has helped Shenyang, Dalian, Chaoyang, Benxi, Anshan, Jinzhou, Huludao and other animal disease control centers for sample pretreatment of African swine fever testing experiments. .

The high-efficiency, stable and safe instrument characteristics of Scientz-48L frozen high-flux tissue grinder well solve the problems of many samples in the detection process and the difficulty in controlling the consistency of the experiment, which has been well received by users.

Product Usage

1. Suitable for grinding and crushing various plant tissues including roots, stems, leaves, flowers, fruits, seeds, etc. 2. Suitable for various animal tissues including brain, heart, lung, stomach, liver, thymus, kidney, intestine Grinding and crushing of lymph nodes, muscles, bones, etc.; 3. Suitable for grinding and crushing of fungi, bacteria and other samples; 4. Grinding and crushing for food and pharmaceutical composition analysis; 5. Suitable for volatile samples including coal and oil Grinding and crushing of samples such as shale and wax products; 6. Suitable for grinding and crushing of plastics, polymers including PE, PS, textiles, resins and the like.

The main technical parameters

1. Time setting: 1 (seconds) - 9999 (seconds)

2. Frequency setting: 10-70Hz, that is, oscillation 300-2100 times/min

3. Rated power: 180W

4. Fixture stroke: 34mm (vertical)

5. Temperature setting: -20 ° C or -30 ° C

6. Electromagnetic lock control opens the door

7. Sample capacity: random standard: aluminum alloy adapter 5ml 12 holes, 2ml 48 holes

     Optional: Aluminum adapter 2ml24 hole

8. Power requirements: 220V single phase AC, <2.5A

Scientz biotechnology company profile

Ningbo scientz biotechnology co.,ltd (stock code: 430685), which is headquartered in the National High-tech Zone of Ningbo. It is a renowned provider of biological sample processing equipment and also a domestic ultrasonic technology application. 

The company currently has four R&D centers in Ningbo and Hangzhou, with dozens of high-level R&D personnel and more than 60 patents.

The company attaches great importance to brand cultivation and market development. In order to better serve customers, the company has more than 30 offices throughout the country. The offices are equipped with sales engineers and after-sales engineers to meet customer needs at any time.  The products are exported to many countries including the United States, Britain, France, Russia, Japan and Korea. A well-known expert in the development of biological sample processing equipment.