Biomaterial Database

Cross-priming amplification targeting the coagulase gene for rapid detection of coagulase-positive Staphylococci. 2015

B Qiao, and J-Y Cui, and L Sun, and S Yang, and Y-L Zhao

School of Chemical Engineering and Technology, Tianjin University, Tianjin, China.

OBJECTIVE To develop and evaluate cross-priming amplification (CPA) combined with immuno-blotting for the detection of coagulase-positive Staphylococci including Staphylococcus aureus. RESULTS Twenty-four sets of cross and detection primers were designed according to four sequences of coagulase gene in Staph. aureus. The most specific primer pair was screened out for the next amplification and interaction. The specificity was evaluated in a total of 53 species of Staph. aureus and non-Staph. aureus. Two red lines indicating positive were always observed on the BioHelix Express strip for 12 subspecies of Staph. aureus. In contrast, only one signal line showing negative results was detected in all of non-Staph. aureus samples. The limit of detection (LOD) of CPA was 3·6 ± 2·7 fg for the genomic DNA, which is about 100 and 10 times sensitive than those of PCR and loop-mediated isothermal amplification respectively. For the pure culture of Staph. aureus and milk powders, the LODs of CPA were about 1·34 CFU per reaction and 5·2 ± 3·7 CFU per 100 g of milk powder respectively. The CPA method was also successfully applied to evaluate the contamination of Staph. aureus in 318 samples of daily food. CONCLUSIONS CPA is a very sensitive and rapid method to detect Staph. aureus by simple laboratory instrument. CONCLUSIONS It is the first report on the application of the CPA with immuno-blotting for detection of coagulase-positive Staphylococci including Staph. aureus.

UI	MeSH Term	Description	Entries
D003030	Coagulase	Enzymes that cause coagulation in plasma by forming a complex with human PROTHROMBIN. Coagulases are produced by certain STAPHYLOCOCCUS and YERSINIA PESTIS. Staphylococci produce two types of coagulase: Staphylocoagulase, a free coagulase that produces true clotting of plasma, and Staphylococcal clumping factor, a bound coagulase in the cell wall that induces clumping of cells in the presence of fibrinogen.	Staphylocoagulase,Staphylococcal Clumping Factor,Clumping Factor (Staphylococcal),Staphylococcus aureus clone pSCa2 of Coagulase,Staphylococcus aureus strain 213 of Coagulase,Staphylococcus aureus strain 8325-4 of Coagulase,Clumping Factor, Staphylococcal,Factor, Staphylococcal Clumping,Staphylococcus aureus strain 8325 4 of Coagulase
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001426	Bacterial Proteins	Proteins found in any species of bacterium.	Bacterial Gene Products,Bacterial Gene Proteins,Gene Products, Bacterial,Bacterial Gene Product,Bacterial Gene Protein,Bacterial Protein,Gene Product, Bacterial,Gene Protein, Bacterial,Gene Proteins, Bacterial,Protein, Bacterial,Proteins, Bacterial
D013203	Staphylococcal Infections	Infections with bacteria of the genus STAPHYLOCOCCUS.	Infections, Staphylococcal,Staphylococcus aureus Infection,Staphylococcal Infection,Staphylococcus aureus Infections
D013210	Staphylococcus	A genus of gram-positive, facultatively anaerobic, coccoid bacteria. Its organisms occur singly, in pairs, and in tetrads and characteristically divide in more than one plane to form irregular clusters. Natural populations of Staphylococcus are found on the skin and mucous membranes of warm-blooded animals. Some species are opportunistic pathogens of humans and animals.
D013211	Staphylococcus aureus	Potentially pathogenic bacteria found in nasal membranes, skin, hair follicles, and perineum of warm-blooded animals. They may cause a wide range of infections and intoxications.
D017931	DNA Primers	Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.	DNA Primer,Oligodeoxyribonucleotide Primer,Oligodeoxyribonucleotide Primers,Oligonucleotide Primer,Oligonucleotide Primers,Primer, DNA,Primer, Oligodeoxyribonucleotide,Primer, Oligonucleotide,Primers, DNA,Primers, Oligodeoxyribonucleotide,Primers, Oligonucleotide
D021141	Nucleic Acid Amplification Techniques	Laboratory techniques that involve the in-vitro synthesis of many copies of DNA or RNA from one original template.	DNA Amplification Technic,DNA Amplification Technique,DNA Amplification Techniques,Nucleic Acid Amplification Technic,Nucleic Acid Amplification Technique,RNA Amplification Technic,RNA Amplification Technique,RNA Amplification Techniques,Amplification Technics, Nucleic Acid,Amplification Techniques, Nucleic Acid,DNA Amplification Technics,Nucleic Acid Amplification Technics,Nucleic Acid Amplification Test,Nucleic Acid Amplification Tests,RNA Amplification Technics,Technics, Nucleic Acid Amplification,Techniques, Nucleic Acid Amplification,Amplification Technic, DNA,Amplification Technic, RNA,Amplification Technics, DNA,Amplification Technics, RNA,Amplification Technique, DNA,Amplification Technique, RNA,Amplification Techniques, DNA,Amplification Techniques, RNA,Technic, DNA Amplification,Technic, RNA Amplification,Technics, DNA Amplification,Technics, RNA Amplification,Technique, DNA Amplification,Technique, RNA Amplification,Techniques, DNA Amplification,Techniques, RNA Amplification