title: A quantitative TaqMan PCR assay for the detection of Mycoplasma suis.
abstract: OBJECTIVE
  To develop a TaqMan probe-based, highly sensitive and specific quantitative PCR (qPCR) assay for the detection and quantification of Mycoplasma suis in the blood of pigs.
  
  
  RESULTS
  Primers and probes specific to Myc. suis 16S rRNA gene were designed. The qPCR assay's specificity, detection limit, intra- and inter-assay variability were evaluated and its performance was compared with a Myc. suis conventional PCR assay (cPCR). Blood of two experimentally infected pigs, 40 Indiana pigs, 40 Brazilian sows and 28 peccaries were tested. The assay detected as few as ten copies of Myc. suis plasmids and was 100-fold more sensitive than the cPCR. No cross-reactivity with nontarget pig mycoplasmas was observed. An average of 1·62 × 10(11) and 2·75 × 10(8) target copies ml(-1) of blood were detected in the acutely and chronically infected pigs, respectively. Three (7·5%) pigs and 32 (80·0%) sows were positive while all peccaries were negative for Myc. suis.
  
  
  CONCLUSIONS
  The developed qPCR assay is highly sensitive and specific for Myc. suis detection and quantification.
  
  
  CONCLUSIONS
  TaqMan qPCR is an accurate and quick test for detection of Myc. suis infected pigs, which can be used on varied instrumentation platforms.
authors.affiliation: Department of Comparative Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: A M S
authors.initials: AM
authors.lastname: Guimaraes
authors.forename: R F C
authors.initials: RF
authors.lastname: Vieira
authors.forename: R
authors.initials: R
authors.lastname: Poletto
authors.forename: R
authors.initials: R
authors.lastname: Vemulapalli
authors.forename: A P
authors.initials: AP
authors.lastname: Santos
authors.forename: W
authors.initials: W
authors.lastname: de Moraes
authors.forename: Z S
authors.initials: ZS
authors.lastname: Cubas
authors.forename: L C
authors.initials: LC
authors.lastname: Santos
authors.forename: J N
authors.initials: JN
authors.lastname: Marchant-Forde
authors.forename: J
authors.initials: J
authors.lastname: Timenetsky
authors.forename: A W
authors.initials: AW
authors.lastname: Biondo
authors.forename: J B
authors.initials: JB
authors.lastname: Messick
mesh_terms: D000818:Animals; D001938:Brazil; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005260:Female; D007196:Indiana; D057230:Limit of Detection; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000235:genetics / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D034421:Sus scrofa / Q000382:microbiology; D013552:Swine / Q000382:microbiology*; D013553:Swine Diseases / Q000097:blood / Q000175:diagnosis*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S