Molecular Diagnostics
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Molecular Diagnostics Tests and Fees Contact Information Scott Hahn, Research Assistant Mike Hays, Technician Sean Smith, Lab Technician |
Tests
PCR is the appropriate test to use for Tritrichomonas foetus diagnosis due to its sensitivity and ability to distinguish between non-pathogenic enteric trichomonads and the pathogenic Tritrichomonas foetus organism.
Follow this link to an instructional video presenting proper Tritrichomonas foetus sampling techniques and appropriate sample handling.
LAD-1000 Pathogenic Leptospira PCR
The purpose of the procedure is to diagnose Leptospira infection or shedding of the organism using polymerase chain reaction technology to amplify a Leptospira genus conserved region of the 23S rDNA (Woo, et al, J. Clin Mic., Dec. 1997, p.3140-3146). The pathogenic Leptospira serovars are then distinguished from saprophytic serovars using a novel FAM-labeled Taqman probe incorporated into a semi-nested PCR detection step on a real time PCR system.
LAD-1010 E. coli O157:H7 Genotyping (eae, STX-1&2) PCR
The purpose of this assay is to better define E. coli O157:H7 isolates for epidemiological purposes by determining the genotyping pattern obtained from separate PCR assays for the entering and effacing (eae) gene, the shiga-like toxin 1 (STX-1) gene and the shiga-like toxin 2 (STX-2) gene.
LAD-1020-1026 Salmonella sp. Enrichment Taqman PCR
Salmonellosis is one of the most common infectious diseases in the world in both humans and animals. Salmonella infections can be manifested in three forms, gastroenteritis, enteric fever and septicemia. Chronic asymptomatic carriers often arise from a population infected with Salmonella serovars and the difficulty in detecting carriers by standard culture techniques makes these carriers a potential source of environmental contamination. We have devised a procedure than combines a short cultivation period in selective media with a PCR Taqman Assay based on Salmonella InvA gene sequence for the identification of Salmonella serovars in culture and clinical specimens.
LAD-1030 Clostridium perfringens genotyping PCR
Clostridial enterotoxemia is an acute, highly fatal condition found in humans, sheep, calves, pigs and horses. It is caused by infection with Clostridium perfringens. Traditionally, production of one or more of four exotoxins (alpha, beta, epsilon and iota toxins) has allowed division of C. perfringens into five toxigenic types. The major diseases caused by these types are listed below.
While not used in determining the classical toxin phenotype, enterotoxin is the principal toxin involved in human food borne illness, and is considered by many to be a virulence attribute in animal strains of C. perfringens. The gene for enterotoxin may be present in any C. perfringens toxin type. Recently, beta2, a novel C. perfringens toxin was identified in strains associated with porcine enteritis. However, a definitive role for this toxin in disease has not been established. Beta2 may be present in any C. perfringens toxin type.
Typing of C. perfringens by toxin neutralization tests in mice or guinea pigs is a traditional part of the diagnosis of clostridial enteritis in domestic animals. For a number of reasons, including cost, lack of availability of standard antitoxin, unexplained variability in results, and humanitarian concerns for animal welfare, in vivo testing is infrequently used at present.
As an alternative to in vivo typing, Dr. Glen Songer at the University of Arizona has developed and evaluated a genotyping system, based upon detection by polymerase chain reaction (PCR) of the exotoxin genes for alpha(cpa), beta(cpb), epsilon(etx), iota(iA)and beta2(cpb2) toxins, and the enterotoxin(cpe) gene.
LAD-1040 - E. coli pilus/toxin PCR multiplex
The E. coli pilus/toxin multiplex PCR detects five pilus and four toxin genes that are commonly identified in enterotoxigenic(ETEC) E. coli of swine, bovine, caprine and ovine species. The test can also be used on E. coli isolates from canine as the STa, STb and SLT2 toxin genes are found in canine ETEC. PCR is performed on single isolates of E. coli cultures from the small intestine or from feces that have been subcultured on blood agar, E-agar or Minca agar.
Pilus Genes
K88 (F4) – Commonly associated with ETEC isolates from porcine
K99 (F5) – Associated with ETEC strains from porcine, bovine and ovine
987P (F6) – Associated with strains from porcine
F41 – Associated with ETEC strains isolated from porcine, bovine and sheep
F107 (F18) – Associated with porcine edema disease and post-weaning diarrhea.
Toxin Genes
LT – Heat labile enterotoxin
STa – Heat stable enterotoxin a
STb – Heat stable enterotoxin b
SLT-2 – Shiga-like toxin 2
LAD-1055 Chlamydia sp. Nested PCR
Chlamydia species are obligate, intracellular bacterial parasites that cause a variety of diseases in mammals and birds. A set of degenerate primers for the Chlamydia genus outer membrane protein 1 (OMP1) have been developed (Kaltenboeck, et al, J. Bacteriology 175:487-502, 1993.) in a nested PCR for the detection of Chlamydia from clinical specimens. Strains of Chlamydophila (abortus, psittaci, felis, caviae, pecorum) and strains of Chlamydia (suis, trachomatis and pneumoniae ) are all detectable with this assay.
LAD-1060 Mycoplasma Species General Nested PCR
Mycoplasmas are the smallest known bacteria and lack a cell wall. They produce and act as a cofactor in respiratory disease, arthritis, conjunctivitis and reproductive problems. The nested PCR assay using commercially available primers published by Harasawa R., et al, Rapid Diagnosis of Mycoplasmas, Edited by Kahane and Adoni, 1993 is used to amplify species characteristic amplicons from the 16S/23S intergenic regions. Outer PCR amplicons can also be used in a restriction fragment length polymorphism (RFLP) assay as described by Lauerman, et al. in his Nucleic Acid Amplification Assays for Diagnosis of Animal Diseases Laboratory Manual.
LAD-1070-1090 Mycoplasma Species Specific PCR
Mycoplasmas are the smallest known bacteria and lack a cell wall. They produce and act as a cofactor in respiratory disease, arthritis, conjunctivitis and reproductive problems. The specific PCR assays use a basic formulation of SYBR Green PCR mix using specific primers for strains of interest published by Lauerman, et al. in his Nucleic Acid Amplification Assays for Diagnosis of Animal Diseases Laboratory Manual. The dissociation temperatures of the resulting amplicons are verified as compared to control strains of Mycoplasma.
LAD-3005 Influenza Universal Taqman RT-PCR
The influenza virus associated with kennel cough respiratory disease in racing greyhounds is a member of the genus Influenza A. Influenza A viruses are known to infect humans, pigs, horses, birds, mink and sea mammals. The natural hosts and source for all influenza A viruses are water birds, especially waterfowl and shorebirds. Most infected greyhounds has clinical disease similar to that seen in horses infected with Equine Influenza A, they exhibit fever and dry hacking cough along with a reluctance to eat and drink. Whereas horses typically recover in two to three weeks, the disease has been known to be fatal to dogs. The Influenza Universal Taqman RT-PCR has been developed to detect all group A influenza viruses from Kansas Racing Commission Kennel Cough swab samples.
LAD-3010 Canine Coronavirus Taqman RT-PCR
Coronaviruses are large, enveloped, positive-stranded RNA viruses. Canine coronavirus is usually responsible for mild, self-limiting infections restricted to the enteric tract. In 2003, a canine group 2 coronavirus was isolate and associated with canine respiratory disease in kennel environments. We have adapted the RT-PCR method of Decaro (N. Decaro et al., J. Virological Methods 119 (2004) 145-150) to monitor Kansas Racing Commission Kennel Cough swab samples for Canine coronavirus group 2.
LAD-3020 Canine Adenovirus PCR
Adenovirus is a linear, double stranded, non-enveloped, DNA virus that is approximately 70-90 nm in diameter. The virus has been associated with 'Kennel Cough' syndrome, a highly contagious tracheobronchitis found in dogs. The disease is found worldwide and will infect a very high percentage of dogs in their lifetime. There are many different agents that contribute to the disease process of tracheobronchitis. The most common are Parainfluenza, Bordetella bronchiseptica, and Mycoplasma. Adenovirus, Reovirus, and Herpes virus are thought to possibly contribute to the disease. Although any one of these organisms can cause symptoms of the disease, the majority of cases are the result of more than one organism. The Canine Adenovirus PCR assay has been developed to detect all Adenovirus from Kansas Racing Commission Kennel Cough Project swab samples.
LAD-3020 Canine Herpes virus PCR
Herpes viruses are enveloped, dsDNA viruses that are approximately 100nm in diameter. Canine Herpes virus is a contagious, often fatal disease of puppies less than 3 weeks of age characterized by a viremia with necrosis and hemorrhage in the liver, kidney and lungs, among other organs. The disease in previously unexposed adults is characterized by mild rhinitis and vaginitis, which may lead to abortion or infertility. The virus has been associated with 'Kennel Cough' syndrome, a highly contagious tracheobronchitis found in dogs. The disease is found worldwide and will infect a very high percentage of dogs in their lifetime. There are many different agents that contribute to the disease process of tracheobronchitis. The most common are Parainfluenza, Bordetella bronchiseptica, and Mycoplasma. Canine Adenovirus, Reovirus, and Herpes virus are thought to possibly contribute to the disease. Although any one of these organisms can cause symptoms of the disease, the majority of cases are the result of more than one organism. The Canine Herpes virus PCR assay has been developed to detect all Herpes virus from Kansas Racing Commission Kennel Cough Project swab samples.