Multiplex PCR Detection Kit for Shiga Toxin-Producing E. Coli Rapid Testing in Food Fluorescent Probe Assays

• Customization: Available
• Specific Usage: For Biological Purpose, For Microbiological
• Content: Standard
• Model NO.: FZ017BF2
• Usage: Laboratory Reagents, Analytical Reagents, Diagnostic Reagents, Teaching Reagents
• Habit Appellation: Chemical Reagent
• Application: Industry, Scientific Research, Health, Environmental Protection, Agriculture
• Sensitivity: Minimum: 100 Cfu/Test
• Shelf Life: 1 Years
• Feature: Easy Operation
• Customized Support: OEM
• Transport Package: Carton
• Specification: 24 tests/box
• Trademark: BHK
• Origin: Guangdong, China
• HS Code: 3821000000

Product Description

Product Description

Introduction

This kit is only suitable for qualitative detection of the type of Shiga toxin produced by pathogenic Escherichia coli (STEC) in
food.
Principle

Based on Real Time PCR technology, primer sets, fluorescent probes and other reagents required for reaction optimization configuration are used for the 16S rRNA gene of Enterobacteriaceae and the Shiga toxin genes stx1 and stx2 of STEC. The nucleic acid of the sample to be tested is added to carry out the amplification reaction. During the amplification process, the fluorescent probe binds to the target gene fragment, which can be decomposed by Taq enzyme and produce a fluorescent signal. At this time, the fluorescent quantitative PCR instrument can recognize the fluorescent signal and draw the corresponding real-time amplification curve according to its intensity change, thereby determining the type of Shiga toxin produced by pathogenic Escherichia coli (STEC). stx1, stx2, and 16S rRNA genes correspond to fluorescent reporter groups HEX, FAM, and Cy5, respectively.

 

How to use?
1. Sample pretreatment 1) Referring to GB 4789.6-2016, use a 1 μL inoculation loop to scrape the suspected colonies to be identified on the nutrient agar plate or slant that have been cultured for 18 h~24 h, suspend them in a sterile centrifuge tube containing 30 μL lysis buffer, fully suspend the bacteria, tap the tube wall to eliminate bubbles, and heat at 99 for 10 min; 2) Centrifuge at 12000 r/min for 15 min. The supernatant is the crude DNA, which can be transferred to a 0.2 mL sterile centrifuge tube and stored for a long time at -20. You can also choose a commercial bacterial genome extraction kit for extraction; 2. Sample addition and reaction 1) Take n PCR reaction tubes as required (n=1 tube of negative control + number of samples to be tested + 1 tube of positive control), take out the premix from the kit, fully melt, vortex and briefly centrifuge, add 20 μL of premix to each tube, and set aside. 2) Add 5 μL of negative control, sample DNA, and positive control to the above n reaction tubes, respectively, with a total reaction volume of 25 μL. Close the tube cap tightly, centrifuge briefly, and immediately perform PCR amplification reaction. 3) The PCR reaction system is 25 μL, take HEX, FAM, and Cy5 detection channels, and collect fluorescence signals at 60°C in reaction stage 2. The specific procedures are as follows:

Reaction stage	Temperature	Time	Signal collection	Number of cycles
1	95°C	30 sec	/	1
2	95°C	5 sec	/	} 40
		60°C	30 sec

Note: Collect fluorescence signals at 60 in reaction stage 2. For multi-channel fluorescence PCR instrument, select fluorescein "HEX", "FAM", and "Cy5" as signal collection channels, select "None" for quenching group, and select "None" for dye correction.

3. Results: In general, the test results can be directly read through the baseline and threshold automatically set by the software. If adjustment is required, it can be adjusted according to the instrument's own conditions (such as noise, etc.) and the different selected fluorescence channels. 1) Quality control: The negative control does not show an obvious S-type amplification curve or the Ct value is greater than 37, and the positive control shows an S-type amplification curve and its Ct value is less than 30. If the positive and negative controls do not meet the above conditions at the same time, the test results are invalid and should be retested or contact product technical support. 2) Result interpretation: (interpretation based on the Ct value obtained by the reaction, see the table below for details:)

Ct value			Result interpretation
≤35			The corresponding gene is positive.
35~37			It is recommended to retest. If the retest result Ct≥37, the corresponding gene is negative, otherwise it is positive.
≥37			The corresponding gene is negative.

Specification

Our service

1.Offering Sample for Testing .
2.R & D team to provide technical support.
3.Provide packaging, labeling OEM customization.

Packing & Delivery

To better ensure the safety of your goods, professional, environmentally friendly, convenient and efficient packaging services will be provided.

Company Profile

 

Guangdong HuanKai Microbial Sci. & Tech. Co.,Ltd. is a national high-tech enterprise under the Guangdong Institute of Microbiology which is attached to Guangdong Academy of Sciences. Since established in 1993, HuanKai has adhered to the strategy of scientific and technological innovation and development, and has formed four product lines including microbiological testing reagents and consumables, digital laboratory instruments, physical and chemical rapid water detecting products, and high-efficiency environmentally friendly cleaning agents and disinfectant, both are with independent intellectual property rights. More than 2, 000 varieties of products are widely applied in mainland China, Hong Kong, Macao, Taiwan and Southeast Asia. Huankai has developed into an important R&D production base for domestic food and other industry.

FAQ

1. Who are we?

HuanKai are based in Guangdong, China, start from 1993,there are total about 301-500 people in HKM. HuanKai has successfully established four major product lines: microbiological testing reagents and consumables, digital laboratory instruments, rapid water testing products for physical and chemical analysis, and eco-friendly cleaning and disinfection solutions. These lines boast over 2,000 different products, each protected by independent intellectual property rights.

2. How can we guarantee quality?
Always a pre-production sample before mass production;
Always final Inspection before shipment;

3.What can you buy from us?
Microbial Detection Product,Peptone&Tryptone,Deydrated Culture Media,Granular Media,Microbial Air Sampler and so on.

4. Why should you buy from us not from other suppliers?
1.With strong R&D capacity and good scientific research conditions,over 30 years of experience 2.One of the largest physicochemical reagent and culture media manufactuer in China follow international standard 3.Stable high quality main raw material from Europe & strictly quality control

5. What services can we provide?
Accepted Payment Currency:USD,EUR,CNY;
Accepted Payment Type: T/T,MoneyGram,PayPal,Western Union,Cash,Escrow;
Language Spoken:English,Chinese