title: Rapid real-time PCR assays for detection of Klebsiella pneumoniae with the rmpA or magA genes associated with the hypermucoviscosity phenotype: screening of nonhuman primates.
abstract: The relationship of mucoviscosity-associated (magA) and/or regulator of mucoid phenotype (rmpA) genes to the Klebsiella pneumoniae hypermucoviscosity (HMV) phenotype has been reported. We previously demonstrated that rmpA+ K. pneumoniae can cause serious disease in African green monkeys and isolated rmpA+ and magA+ HMV K. pneumoniae from other species of non-human primates. To rapidly screen African green monkeys/non-human primates for these infections, we developed three real-time PCR assays. The first was K. pneumoniae-specific, targeting the khe gene, while the others targeted rmpA and magA. Primer Express 2 was used with the three K. pneumoniae genes to generate sequence-specific TaqMan/TaqMan-Minor Groove Binder assays. Oral/rectal swabs and necropsy samples were collected; swabs were used for routine culture and DNA extraction. K. pneumoniae colonies were identified on the Vitek 2 with DNA tested using the K. pneumoniae-specific assays. Testing of 45 African green monkeys resulted in 19 khe+ samples from 14 animals with none positive for either rmpA or magA. Of these 19 khe+ samples, five were culture-positive, but none were HMV "string test"-positive. Subsequent testing of 307 non-human primates resulted in 64 HMV K. pneumoniae isolates of which 42 were rmpA+ and 15 were magA+. Non-human primate testing at the U.S. Army Medical Research Institute of Infectious Diseases demonstrated the ability to screen both live and necropsied animals for K. pneumoniae by culture and real-time PCR to determine HMV genotype.
authors: Diagnostic Systems Division, Veterinary Pathology, U.S. Army Medical Research Institute, of Infectious Diseases, Fort Detrick, Frederick, Maryland, USA.
authors: Laurie J
authors: LJ
authors: Hartman
authors: Edward B
authors: EB
authors: Selby
authors: Chris A
authors: CA
authors: Whitehouse
authors: Susan R
authors: SR
authors: Coyne
authors: James G
authors: JG
authors: Jaissle
authors: Nancy A
authors: NA
authors: Twenhafel
authors: Robin L
authors: RL
authors: Burke
authors: David A
authors: DA
authors: Kulesh
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics*; D002522:Chlorocebus aethiops; D007710:Klebsiella Infections / Q000175:diagnosis* / Q000382:microbiology*; D007711:Klebsiella pneumoniae / Q000235:genetics* / Q000302:isolation & purification; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D011323:Primates; D014783:Viscosity
chemical_list: D001426:Bacterial Proteins; C501254:MagA protein, Klebsiella pneumoniae; C512267:RmpA protein, Klebsiella pneumoniae