Diversity ofMycoplasma hyopneumoniaein pig farms revealed by direct molecular typing of clinical material

Abstract

-Mycoplasma hyopneumoniae is the etiological agent of enzootic pneumonia in swine.Various reports indicate that different strains are circulating in the swine population. We investigated the variety of M. hyopneumoniae strains by a newly developed genetic typing method based on the polyserine repeat motif of the LppS homolog P146. PCR amplification using M. hyopneumoniae specific, conserved primers flanking the region encoding the repeat motif, followed by sequencing and cluster analysis was carried out. The study included strains isolated from different geographic regions as well as lysates from lung swabs from a series of pig farms in Switzerland. High diversity of M. hyopneumoniae was observed but farms being in close geographic or operative contact generally seemed to be affected by the same strains. Moreover, analysis of multiple samples from single pig farms indicated that these harbored the same, farm-specific strain. The results indicate that multiple strains of M. hyopneumoniae are found in the swine population but that specific strains or clones are responsible for local outbreaks. The method presented is a highly reproducible epidemiologic tool allowing direct typing of M. hyopneumoniae from clinical material without prior isolation and cultivation of strains. enzootic pneumonia / epidemiology / LppS / P146

Multiple-Locus Variable-Number Tandem-Repeat Analysis Is a Suitable Tool for Differentiation of Mycoplasma hyopneumoniae Strains without Cultivation

“…Thus, molecular typing methods able to differentiate strains are valuable both for epidemiological studies and experimental trials. Currently, available methods are relatively expensive and labor-intensive, as they require either cultivation of this fastidious organism or sequencing (6,7,11). Using MLVA (multiple-locus VNTR [variable-number tandem-repeat] analysis), we aimed to develop a quick and relatively inexpensive assay able to differentiate strains in samples from the respiratory tracts of pigs without prior cultivation.…”

Real-Time PCR Assays To Address Genetic Diversity among Strains ofMycoplasma hyopneumoniae

“…A number of PCR assays have been developed and reported to be both sensitive and specific for M. hyopneumoniae (4,6,11,19,20,22,24,26). As a result, this technique is now among the most widely used for detection of M. hyopneumoniae in pigs.While the development of PCR assays has greatly enhanced our ability to detect M. hyopneumoniae, the genetic variability of the organism (1,5,12,14,15,21) can affect detection by PCR. It is not known what effect this heterogeneity has on the sensitivities of the different assays.…”

“…While the development of PCR assays has greatly enhanced our ability to detect M. hyopneumoniae, the genetic variability of the organism (1,5,12,14,15,21) can affect detection by PCR. It is not known what effect this heterogeneity has on the sensitivities of the different assays.…”

Survey Of Infectious Agents Associated With Porcine Respiratory Disease Complex (PRDC) In Serbian Swine Herds Using Polymerase Chain Reaction (PCR) Detection

“…The lysed samples were processed with a DNA extraction kit (QIA extraction DNA kit, Qiagen) according to the manufacturer's instructions. All samples were tested by PCR using HotStarTaq Master Mix Kit (Qiagen), and primers described for DNA detection of PCV2 [11], Mhp [12], Pm [13], App [14], Hps [15], Ssuis [16] and Apyo [17]. After suspension, the samples were processed with a RNA extraction kit (Qiagen -Rneasy MiniKit) according to th e manufacturer's instructions.…”