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The results of the multiplex RT-PCR were consistent with those of other diagnoses, such as uniplex RT-PCR, to detect each of the pathogens.
Following in silico screening for candidate target genes representing each of the pathogens, an optimised multiplex PCR assay was established using three primer sets, cytob1, MAR1bB2 and bovar2A, for amplification of genomic DNA of T. annulata, A. marginale and B. bovis respectively.
We assessed whether each of the pathogens of interest were enzootic or epizootic in the YNP canid community, and whether pathogen exposure varied by region of the park in relation to canid density.
For each of the pathogens analysed, downloading was started from the HAMAP tool under the ExPASy web server of all protein sequences whose annotation included the following keywords: Outer, membrane, lipoprotein, adhesin, surface, secreted or exposed.
The orders had different levels of activity against each of the pathogens.
In general, the genes showed similar expression patterns in response to each of the pathogens.
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We generated comprehensive protein interaction maps by performing a ten-fold coverage of the coding capacity of each of the pathogen genomes.
The whole procedure was repeated separately for each of the pathogen isolates.
To facilitate comparison of the activities of different orders against different pathogens classes, the mean MIC's of the orders against each of the pathogen classes were ranked from high activity (low number) to low activity (high number) (Table 5).
We assumed a constant syndrome level attributable to factors other than the respiratory pathogens and constant scaling factors for each of the lagged pathogens.
Thus, in PERCH we are aiming to develop an innovative statistical approach to model the attributable risk of pneumonia for each of these pathogens that can handle the complexity of the PERCH data set and address the correlation among pathogen infections in the presence of possible measurement errors.
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