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590548 genesig Real-Time PCR Detection Kit for Invasion protein (invA) gene (All pathogenic Salmonella species) Standard

Specifications
Kit Type
PCR
Tests
150
Sensitivity
10-100 copies
Detection Method
Fluorescent
Sample Matrix
Unlimited
Shipping Temp
RT
Storage Temp
-20°C

Salmonella spp. are members of the family Enterobacteriaceae. They are Gram-negative, facultatively anaerobic, flagellated, rod-shaped organisms. They are approximately 0.7-1.5µm in diameter and 2-5um in length and responsible for a large number of cases of foodborne illness throughout the world. Salmonella have circular DNA genomes with a mean length of approximately 4530 kb, although this can vary by up 1000 kb. Salmonella classification is extremely complex, however, the genus is divided into two species: S. enterica and S.bongori. S. enterica is then itself divided into 6 biochemically distinct subspecies and the Salmonella genus is further classified into serovars (serotypes) based on the lipopolysaccharide (O), flagella protein (H), and sometimes the capsular (VI) antigens. There are more than 2500 known serovars and within a serovar there may be strains that differ in virulence. Salmonella are mainly transmitted by the faecal-oral route. They are carried asymptomatically in the intestines or gall bladder of many animals, being continuously or intermittently shed in the faeces. Humans can become infected if they do not wash their hands after contact with infected animals or animal feces. In such instances the bacteria adhere to and enter the cells of the intestinal epithelium. The toxins produced by the bacteria can damage and kill the cells that line the intestines, which results in intestinal fluid loss. The bacteria can survive for weeks in a dry environment and far longer in water thus they are frequently present in polluted waters. Salmonella can also be carried latently in the mesenteric lymph nodes or tonsils; these bacteria are not shed, but can become reactivated after stress or immunosuppression. In addition, fomites and vectors can spread Salmonella and vertical transmission occurs in birds, with contamination of the vitalize membrane, albumen and possibly the yolk of eggs. Salmonella spp. can also be transmitted in utero in mammals. There are two different disease conditions that are distinct to salmonellosis; gastroenteritis and enteric typhoid fever. The gastroenteritis is a nonsystemic infection of the intestinal tract and regional lymph nodes that gives rise to headache, muscle aches, diarrhea, vomiting, abdominal cramping, chills, fever, nausea and dehydration. In contrast, the enteric typhoid fever is a systemic disease in which the microorganism replicates within the cells of the reticuloendothelial system. The symptoms usually appear 6 to 72 hours after ingesting contaminated food although individuals can be infected with the bacteria without having symptoms. Those with and without symptoms shed the bacteria in their stool and it is important that personal hygiene be maintained at all times.

The genesig® Kit for for All pathogenic Salmonella species (Salmonella_invA) genomes is designed for the in vitro quantification of Salmonella_invA genomes. The kit is designed to have a broad detection profile. Specifically, the primers represent 100% homology with over 95% of the NCBI database reference sequences available at the time of design.
Product features
• Exceptional value for money • Rapid detection of all clinically relevant subtypes • Positive copy no. standard curve for quantification • Highly specific detection profile • High priming efficiency • Broad dynamic detection range (>6 logs) • Sensitive to < 100 copies of target • Accurate controls to confirm findings
Suitable Sample Material
All kinds of sample material suited for PCR amplification can be used. Please ensure the samples are suitable in terms of purity, concentration, and DNA integrity. Always run at least one negative control with the samples. To prepare a negative control, replace the template DNA sample with RNAse/DNAse free water.
Dynamic Range of Test
Under optimal PCR conditions genesig Salmonella_invA detection kits have very high priming efficiencies of >95% and can detect less than 100 copies of target template.
Product Manual
https://www.genesig.com/assets/files/salmonella_inva_std.pdf
Test Principle
A Salmonella_invA specific primer and probe mix is provided and this can be detected through the FAM channel. The primer and probe mix provided exploits the so-called TaqMan® principle. During PCR amplification, forward and reverse primers hybridize to the Salmonella_invA DNA. A fluorogenic probe is included in the same reaction mixture which consists of a DNA probe labeled with a 5`-dye and a 3`-quencher. During PCR amplification, the probe is cleaved and the reporter dye and quencher are separated. The resulting increase in fluorescence can be detected on a range of qPCR platforms.
Standard Kit Components
• Salmonella_invA specific primer/probe mix (150 reactions BROWN); FAM labelled • Salmonella_invA positive control template (for Standard curve RED) • RNAse/DNAse free water (WHITE) for resuspension of primer/probe mixes • Template preparation buffer (YELLOW) for resuspension of positive control template and standard curve preparation
Reagents and Equipment to be Supplied by the User
• Real-Time PCR Instrument • DNA extraction kit This kit is recommended for use with genesig EASY DNA/RNA Extraction kit. However, it is designed to work well with all processes that yield high quality DNA with minimal PCR inhibitors. • oasigTM Lyophilised or PrecisionPLUSTM 2 x qPCR Mastermix This kit is designed to work well with all commercially available Mastermixes. However, we recommend the use of oasigTM or PrecisionPLUSTM 2x qPCR MasterMix. • Pipettors and Tips • Vortex and centrifuge • Thin walled 1.5 ml PCR reaction tubes
Kit Storage and Stability
This kit is stable at room temperature but should be stored at --20°C on arrival. Once the lyophilised components have been re-suspended they should not be exposed to temperatures above -20°C for longer than 30 minutes at a time and unnecessary repeated freeze/thawing should be avoided. The kit is stable for six months from the date of resuspension under these circumstances. If a standard curve dilution series is prepared this can be stored frozen for an extended period. If you see any degradation in this serial dilution a fresh standard curve can be prepared from the positive control. We donot recommend using the kit after the expiry date stated on the pack.
Instrumation Requirements
Designed for use with real-time PCR instruments. . For use in genesig® q16, look for the “Easy” nomenclature.
About The genesig® KITS
genesig® kits are sold for research use only and are not licensed for diagnostic procedures. Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative Real-Time PCR kits for the detection and simultaneous quantification of numerous significant pathogens. A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented Real-Time PCR expertise in design and validation ensure the best possible kit. 
Details of the target and priming specificity are included in the individual handbooks above. Packaged, optimized and ready to use.
DNA/RNA Extraction
genesig® Easy DNA/RNA extraction kits are a simple to use, low cost and powerful extraction technology suitable for virtually all sample types.
Important Note
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Toxicity and Hazards
 All products should be handled by qualified personnel only, trained in laboratory procedures.
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