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.
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.
Hartman
Laurie J
LJ
Diagnostic Systems Division, Veterinary Pathology, U.S. Army Medical Research Institute, of Infectious Diseases, Fort Detrick, Frederick, Maryland, USA.
Selby
Edward B
EB
Whitehouse
Chris A
CA
Coyne
Susan R
SR
Jaissle
James G
JG
Twenhafel
Nancy A
NA
Burke
Robin L
RL
Kulesh
David A
DA
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
D001426:Bacterial Proteins; C501254:MagA protein, Klebsiella pneumoniae; C512267:RmpA protein, Klebsiella pneumoniae