Comparative Analysis of Indirect ELISA and Real Time PCR for the Detection of Anaplasma Marginale in Buffalo, Cattle and Sheep in District Peshawar and Lakki Marwat, Pakistan

The current study was conducted to investigate the prevalence of BVD virus in Basrah and Nassirya city by using ELISA and RT-PCR. Two hundreds and eighty two samples of non vaccinated cattle sera samples collected from two regions of Iraq (188 samples from Nassirya city and 92 samples from Basrah city). Samples tested by Enzyme Linked Immunosorbent Assay (ELISA) antigen capture. Positive results were 20 samples ( 8 sample in Thi-Qar and 12 positive samples from Basrah). All samples submitted to indirect ELISA(IDEXX HerdCheck ELISA )for detect BVDV antibodies .Genotyping of all 20 positive samples to antigen detection were tested by Real time PCR, using Cador BVDV ½ kit, after extraction of virus RNA by QIAamp mini kit. The results revealed that there were 20 positive sample according to direct ELISA(Ag detection), while 66 sample were positive to indirect ELISA, as well as, the result of RT-PCR showed that there were two sample positive to BVDV type-1 (one sample form each city).Key words: BVDV, Genotype, ELISA, Iraq, Real time PCR.

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Evaluation of the ability of cholera vibrios to form a biofilm on the surface of the chitinous shell of a crayfish by real-time PCR

Journal of microbiology epidemiology immunobiology ◽

10.36233/0372-9311-99 ◽

2021 ◽

Vol 98 (4) ◽

pp. 434-439

Author(s):

E. A. Menshikova ◽

E. M. Kurbatova ◽

S. O. Vodopyanov ◽

R. V. Pisanov ◽

S. V. Titova

Keyword(s):

Comparative Analysis ◽

Vibrio Cholerae ◽

Real Time ◽

Real Time Pcr ◽

Biofilm Formation ◽

Ecological Niche ◽

Natural Ecosystems ◽

Astacus Astacus

Introduction. Most of the bacteria exist in natural ecosystems not in the form of free floating cells; but in the form of biofilms attached to the substrate. One of the most ecologically important substrates is chitin. Vibrio cholerae; like most members of the Vibrionaceae family; has a chitinolytic complex and can degrade chitin. The ability of V. cholerae to form a biofilm on chitinous substrates can explain the mechanism of the formation of an ecological niche for the preservation and transfer of the pathogen to new regions with the likelihood of the formation of new foci of cholera.Aim — to determine the ability of V. cholerae to form a biofilm on the chitinous shell of crayfish (Astacus astacus) by means of real-time PCR.Materials and methods. A comparative analysis of the timing of biofilm formation by V. cholerae of different serogroups and toxigenicity was carried out.Results. In the course of the study; it was found that cholera vibrios were shown to be capable of forming a biofilm regardless the serogroup and toxigenicity. However; toxigenic tcpA+ strains have a higher intensity of biofilm formation than nontoxigenic ones; in which the tcpA gene is absent.

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Correction: Comparative Analysis of Gene Expression Level by Quantitative Real-Time PCR Has Limited Application in Objects with Different Morphology

PLoS ONE ◽

10.1371/annotation/fe0ad7f2-2921-4b13-903a-6eba19e8294b ◽

2012 ◽

Vol 7 (8) ◽

Cited By ~ 1

Author(s):

Natalia V. Demidenko ◽

Aleksey A. Penin

Keyword(s):

Gene Expression ◽

Comparative Analysis ◽

Real Time ◽

Real Time Pcr ◽

Gene Expression Level ◽

Expression Level ◽

Quantitative Real Time Pcr ◽

Limited Application

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Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction) based on msp5 gene

Pesquisa Veterinária Brasileira ◽

10.1590/s0100-736x2014000100005 ◽

2014 ◽

Vol 34 (1) ◽

pp. 29-33 ◽

Cited By ~ 4

Author(s):

Gisele M. Bacanelli ◽

Carlos A. N. Ramos ◽

Flábio R. Araújo

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Real Time Pcr ◽

Anaplasma Marginale ◽

Diagnostic Methods ◽

Molecular Diagnostic ◽

Chain Reaction ◽

The Real ◽

End Point ◽

Polymerase Chain

The rickettsia Anaplasma marginale is considered the main agent of bovine anaplasmosis. Due the nonspecific clinical signs of the anaplasmosis, the diagnosis of infection depends of laboratory confirmation. In recent years, molecular diagnostic methods have been used to detect A. marginale in cattle. However, the existence of a large number of assays of different sensitivity and cost makes the choice of an appropriate test difficult. In the present study, a real-time Polymerase Chain Reaction (PCR) based on the msp5 target gene was quantitatively assessed and compared to an end point PCR. Both reactions were subjected to sensitivity and specificity evaluation using plasmid DNA and samples from cattle experimentally infected with A. marginale. A comparative field trial of the tests was carried out using samples of cattle from a stable enzootic area for A. marginale. The real-time PCR showed a higher sensitivity than the end point PCR. This reaction (i.e. real-time PCR) was able to detect one copy of the msp5 gene in 100 ηg of plasmidial DNA, and more than 80% of its results were positive among experimentally infected animals seven days after infection. In addition, based on in silico analysis, the real-time PCR evaluated in the present study appears to be useful for the detection of A. ovis.

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PSVI-41 The comparative analysis of vaginal flora of high-productive cows in the postpartum period by Real-time PCR

Journal of Animal Science ◽

10.1093/jas/skz258.411 ◽

2019 ◽

Vol 97 (Supplement_3) ◽

pp. 199-200

Author(s):

Elena A Korochkina ◽

Andrey Nechaev ◽

Anatoly Stekolnikov ◽

Anatoliy Nezhdanov

Keyword(s):

Comparative Analysis ◽

Real Time ◽

Real Time Pcr ◽

Postpartum Period ◽

Vaginal Discharge ◽

Bacterial Count ◽

Vaginal Flora ◽

High Count ◽

Yeast Fungi ◽

Apparently Healthy

Abstract The aim of the study was the comparative analysis of species composition of vaginal bugs of high-productive cows (n = 7) in the postpartum period by Real-time PCR. The material were vaginal discharge of cows (four of them had acute pyogenic endometritis, another was apparently healthy). The vaginal discharge of sick cows contained the low count of bacteria Megasphaera spp., Veillonella spp., Dialister sрp., Lactobacillus spp. and streptococcus. The evaluation of bacterial count of Lachnobacterium spp., Clostridium spp., actinomycetes Atopobium spp., bacteria Staphylococcus spp., and yeast fungi Candida spр. did not demonstrate statistically different between groups. Probably this bacteria were not the etiological origin of postpartum endometritis of cows. However, vaginal discharge of cows with diagnosis acute pyogenic endometritis had high count of bacteria of Fusobacteriaceae (6,0±3,8 and 5,3±2,6 lg genomes/g), Enterobacteriaceae (6,4±4,1 and 5,1±3,0 lg genomes/g), bacteroides of Prevotella spp. and Porphyromonas spp. (7,4±5,2 and 6,4±4,4 lg genomes/g), actinomycetes Mobiluncus spp. and Corynebacterium spp. (5,1±2,5 and 4,7±1,9 lg genomes/g), Peptostreptococcus spp. and Eubacterium spp., in comparison with apparently healthy cows. The presence of this bacteria in the vaginal discharge of apparently healthy cows suggestive that this bacteria are the permanent vaginal flora. Thus, comparative analysis of the dysbiotic sort of vaginal flora of cows in postpartum period demonstrated that vaginal discharge of cows with diagnosis acute pyogenic endometritis consists the high count of bacteria of potentially pathogenic and pathogenic groups (Fusobacteriaceae, Enterobacteriaceae, Prevotella spp., Porphyromonas spp., Mobiluncus spp., Peptostreptococcus spp., Eubacterium spp.).

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