The use of oral fluids and sock samples for monitoring key pathogens in pig populations for surveillance purposes.

Despite the prevalence of co-infections and the association of over 50 viral and 46 bacterial pathogens with pig diseases, little is known about their simultaneous occurrence, particularly in commercial pig farming environments where health programs are in place. To address this knowledge gap, this study aimed to evaluate the pathogen threshold of respiratory and enteric pathogens in pig herds using the Pork MultiPath™ (PMP1 and PMP2, respiratory and enteric respectively) technology, which detects multiple pathogens simultaneously in a single reaction with high sensitivity and specificity. In this study the most prevalent respiratory pathogens, Mycoplasma hyrohinis, Pasteurella multocida, and Haemophilus parasuis detected by PMP1 were effectively controlled during the nursery stage through strategic treatment with tiamulin.

A comparison of two informative SNP-based strategies for typing Pseudomonas aeruginosa isolates from patients with cystic fibrosis.

Background: Molecular typing is integral for identifying Pseudomonas aeruginosa strains that may be shared between patients with cystic fibrosis (CF). We conducted a side-by-side comparison of two P. aeruginosa genotyping methods utilising informative-single nucleotide polymorphism (SNP) methods; one targeting 10 P.

High-throughput single-nucleotide polymorphism-based typing of shared Pseudomonas aeruginosa strains in cystic fibrosis patients using the Sequenom iPLEX platform.

Shared strains of Pseudomonas aeruginosa are now well recognized in people with cystic fibrosis (CF), and suitable P. aeruginosa laboratory typing tools are pivotal to understanding their clinical significance and guiding infection control policies in CF clinics. We therefore compared a single-nucleotide polymorphism (SNP)-based typing method using Sequenom iPLEX matrix-assisted laser desorption ionization with time-of-flight mass spectrometry (MALDI-TOF MS) with typing methods used routinely by our laboratory.

Cross-platform array screening identifies COL1A2, THBS1, TNFRSF10D and UCHL1 as genes frequently silenced by methylation in melanoma.

Epigenetic regulation of tumor suppressor genes (TSGs) has been shown to play a central role in melanomagenesis. By integrating gene expression and methylation array analysis we identified novel candidate genes frequently methylated in melanoma. We validated the methylation status of the most promising genes using highly sensitive Sequenom Epityper assays in a large panel of melanoma cell lines and resected melanomas, and compared the findings with those from cultured melanocytes.

Multilocus sequence typing of Streptococcus pneumoniae by use of mass spectrometry.

Multilocus sequence typing (MLST) is an important tool for the global surveillance of bacterial pathogens that is performed by comparing the sequences of designated housekeeping genes. We developed and tested a novel mass spectrometry-based method for MLST of Streptococcus pneumoniae. PCR amplicons were subjected to in vitro transcription and base-specific cleavage, followed by analysis of the resultant fragments by using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS).

In vitro manipulation of mammalian preimplantation embryos can alter transcript abundance of histone variants and associated factors.

Manipulation of mammalian embryos and gametes in vitro reduces viability. Specific causes for these reductions are still largely undetermined. Accumulating evidence suggests that survival rates and developmental competency may be reduced following disruptions in the epigenetic regulation of gene expression.

Expression of genes coding for histone variants and histone-associated proteins in pluripotent stem cells and mouse preimplantation embryos.

The histone code is an epigenetic regulatory system thought to play a crucial role in cellular events such as development, differentiation and in the maintenance of pluripotency. In order to gain an insight into the role variant histones may play during mammalian development; we studied gene expression of histone variants and remodelling enzymes in mouse embryonic stem (ES) cells and during mouse preimplantation development. Using quantitative reverse-transcription PCR (qRT-PCR) we document the gene expression pattern of 12 histone variants and 2 of their associated remodelling enzymes in undifferentiated ES cells and during preimplantation embryo development.

Gene expression profiling of porcine peripheral blood leukocytes after infection with Actinobacillus pleuropneumoniae.

The gene expression profile of peripheral blood leukocytes (PBL) from extreme performing pigs after infection with Actinobacillus pleuropneumoniae was analysed using a custom complementary DNA (cDNA) microarray and quantitative reverse transcription-PCR (qRT-PCR). Four high performing animals with low disease-score (HP), three low performing animals with high disease-score (LP) and one medium performing animal with medium disease-score (MP) were selected for microarray profiling. PBL RNA from these eight pigs collected before and at 24h after APP infection, was examined.

Universal reference method for real-time PCR gene expression analysis of preimplantation embryos.

Real-time PCR expression profiling in individual preimplantation embryos poses two main challenges. First, the amount of RNA from blastocysts (between 100 and 200 cells) is too small to quantify, and secondly, a reference gene with stable expression across preimplantation embryos produced by different reproductive technologies is required. We have developed a method using RNA and DNA spikes that allows for accurate normalization of gene expression without the use of an internal housekeeping gene in preimplantation blastocysts.