Biomaterial Database

Verification of an Edwardsiella ictaluri-specific diagnostic PCR. 2010

M L Williams, and M L Lawrence

College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762, USA.

OBJECTIVE To verify the specificity of a PCR assay for the identification and diagnosis of Edwardsiella ictaluri. RESULTS An Edwardsiella ictaluri-specific PCR assay was developed utilizing two features of the ribosomal DNA gene clusters. The first feature is the presence of two ribosomal gene clusters located in tandem to one another (the inter-ribosomal spacer, IRS). This characteristic is present in the Edwardsiella genus but absent in the other sequenced members of the Enterobacteriaceae. The second feature is the presence of an intervening sequence (IVS) in the 23S rRNA gene of Edw. ictaluri. To verify the specificity of this assay, we tested genomic DNA from a variety of bacterial species. The IVS/IRS PCR assay results in an c. 2000-bp product from all Edw. ictaluri isolates tested, but not from any other species including Edwardsiella tarda. CONCLUSIONS The IVS/IRS PCR assay is highly specific for Edw. ictaluri and useful as a tool for identifying this pathogen. CONCLUSIONS This research verifies the specificity of PCR-based assay for Edw. Ictaluri, and we describe this assay as a highly versatile diagnostic tool for its identification.

UI	MeSH Term	Description	Entries
D002397	Catfishes	Common name of the order Siluriformes. This order contains many families and over 2,000 species, including venomous species. Heteropneustes and Plotosus genera have dangerous stings and are aggressive. Most species are passive stingers.	Eremophilus mutisii,Heteropneustes,Plotosus,Siluriformes,Arius,Catfish,Colombian Catfish,Catfish, Colombian
D004269	DNA, Bacterial	Deoxyribonucleic acid that makes up the genetic material of bacteria.	Bacterial DNA
D005393	Fish Diseases	Diseases of freshwater, marine, hatchery or aquarium fish. This term includes diseases of both teleosts (true fish) and elasmobranchs (sharks, rays and skates).	Disease, Fish,Diseases, Fish,Fish Disease
D005810	Multigene Family	A set of genes descended by duplication and variation from some ancestral gene. Such genes may be clustered together on the same chromosome or dispersed on different chromosomes. Examples of multigene families include those that encode the hemoglobins, immunoglobulins, histocompatibility antigens, actins, tubulins, keratins, collagens, heat shock proteins, salivary glue proteins, chorion proteins, cuticle proteins, yolk proteins, and phaseolins, as well as histones, ribosomal RNA, and transfer RNA genes. The latter three are examples of reiterated genes, where hundreds of identical genes are present in a tandem array. (King & Stanfield, A Dictionary of Genetics, 4th ed)	Gene Clusters,Genes, Reiterated,Cluster, Gene,Clusters, Gene,Families, Multigene,Family, Multigene,Gene Cluster,Gene, Reiterated,Multigene Families,Reiterated Gene,Reiterated Genes
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D012329	RNA, Bacterial	Ribonucleic acid in bacteria having regulatory and catalytic roles as well as involvement in protein synthesis.	Bacterial RNA
D012338	RNA, Ribosomal, 23S	Constituent of 50S subunit of prokaryotic ribosomes containing about 3200 nucleotides. 23S rRNA is involved in the initiation of polypeptide synthesis.	23S Ribosomal RNA,23S rRNA,RNA, 23S Ribosomal,Ribosomal RNA, 23S,rRNA, 23S
D012680	Sensitivity and Specificity	Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)	Specificity,Sensitivity,Specificity and Sensitivity
D013045	Species Specificity	The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.	Species Specificities,Specificities, Species,Specificity, Species
D016133	Polymerase Chain Reaction	In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.	Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain