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Basic Info.
Sensitivity
Mimimum:100 Cfu/Test
Feature
Easy Operation
Customized Support
OEM
Transport Package
Carton
Specification
24tests
Trademark
BHK
Origin
Guangdong, China
Product Description
Introduction
This kit is suitable for the detection of Legionella pneumophila in drinking water, non-drinking water, and environmental samples
in public places. Principle
in public places. Principle
Based on Real Time PCR technology, primers and fluorescent probes are designed for the Legionella pneumophila-specific gene mip and the reagent components required for the reaction are optimized. The amplification reaction can be carried out by adding the nucleic acid of the sample to be tested. During the amplification process, the fluorescent probe binds to the target gene fragment, which can be decomposed by the 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 changes, thereby determining whether Legionella pneumophila is detected.
How to use?
1. Sample pretreatment 1) Preparation of sample enrichment solution template DNA: a) The sampling process refers to the national standard "GB/T 18204.3-2013 Public Place Hygiene Inspection Method Part 3: Air Microorganisms", or the method established by each laboratory. After sampling, it can be inoculated into 100 mL GVPC liquid culture medium, placed in a carbon dioxide incubator at 36 ±1 for culture, and enriched for 18~24 hours; b) Aspirate 1 mL of enrichment solution into a 1.5 mL sterile centrifuge tube, centrifuge at 6000 r/min for 5 min, and completely remove the supernatant; c) Add 30 μL lysis solution, fully suspend the bacteria, tap the tube wall to eliminate bubbles, and heat at 99 for 10 min; d) 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 can be stored for a long time at -20. 2) Preparation of template DNA of suspicious colonies: Pick suspicious colonies and fully suspend them in a sterile centrifuge tube pre-added with 30 μL lysis solution, and then follow the above steps c) and d). 2. Sample addition and reaction 1) Take n PCR reaction tubes as required (n = number of samples to be tested + 1 tube of negative control + 1 tube of positive control), take out the premix from the kit, fully melt, vortex and briefly centrifuge, and add 20 μL of premix to each of the above PCR tubes. 2) Add 5 μL of negative control, sample DNA to be tested, and positive control to the above n reaction tubes in order, cover the tube cap immediately after adding the sample, briefly centrifuge, and immediately perform PCR amplification reaction. 3) The PCR reaction system is 25 μL, set the FAM detection channel to read, and collect the fluorescence signal at 60°C in reaction stage 2. The specific procedure is as follows:
1. Sample pretreatment 1) Preparation of sample enrichment solution template DNA: a) The sampling process refers to the national standard "GB/T 18204.3-2013 Public Place Hygiene Inspection Method Part 3: Air Microorganisms", or the method established by each laboratory. After sampling, it can be inoculated into 100 mL GVPC liquid culture medium, placed in a carbon dioxide incubator at 36 ±1 for culture, and enriched for 18~24 hours; b) Aspirate 1 mL of enrichment solution into a 1.5 mL sterile centrifuge tube, centrifuge at 6000 r/min for 5 min, and completely remove the supernatant; c) Add 30 μL lysis solution, fully suspend the bacteria, tap the tube wall to eliminate bubbles, and heat at 99 for 10 min; d) 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 can be stored for a long time at -20. 2) Preparation of template DNA of suspicious colonies: Pick suspicious colonies and fully suspend them in a sterile centrifuge tube pre-added with 30 μL lysis solution, and then follow the above steps c) and d). 2. Sample addition and reaction 1) Take n PCR reaction tubes as required (n = number of samples to be tested + 1 tube of negative control + 1 tube of positive control), take out the premix from the kit, fully melt, vortex and briefly centrifuge, and add 20 μL of premix to each of the above PCR tubes. 2) Add 5 μL of negative control, sample DNA to be tested, and positive control to the above n reaction tubes in order, cover the tube cap immediately after adding the sample, briefly centrifuge, and immediately perform PCR amplification reaction. 3) The PCR reaction system is 25 μL, set the FAM detection channel to read, and collect the fluorescence signal at 60°C in reaction stage 2. The specific procedure is 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: For ABI series fluorescence quantitative PCR instruments, ROX does not need to be added to the reaction system, and when setting the software, select "None" at both "Passive Reference" and "Quencher".
3. Results: In general, the test results can be directly read through the baseline and threshold automatically set by the software. If adjustments are required, they can be made according to the instrument's own conditions (such as noise, etc.) and the different fluorescent channels selected. 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 contacted with 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 | | Legionella pneumophila is positive. | | | |||
| 35~37 | | It is recommended to retest. If the result Ct is ≥37, Legionella pneumophila is negative, otherwise it is positive for Legionella pneumophila. | | | |||
| ≥37 | | Legionella pneumophila is negative. | | | |||
Our service
1.Offering Sample for Testing .
2.R & D team to provide technical support.
3.Provide packaging, labeling OEM customization.
2.R & D team to provide technical support.
3.Provide packaging, labeling OEM customization.
To better ensure the safety of your goods, professional, environmentally friendly, convenient and efficient packaging services will be provided.
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.
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