scholarly journals Investigation of the presence of slime production, VanA gene and antiseptic resistance genes in Staphylococci isolated from bovine mastitis in Algeria

2020 ◽  
Vol 52 (1) ◽  
Author(s):  
Radhwane Saidi ◽  
Zafer Cantekin ◽  
Nora Mimoune ◽  
Yasar Ergun ◽  
Hasan Solmaz ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Resistance Genes ◽  
Detection Rate ◽  
Bovine Mastitis ◽  
Mammary Glands ◽  
Chain Reaction ◽  
Vana Gene ◽  
Slime Production ◽  
Pcr Method ◽  
Polymerase Chain

Staphylococcus strains are frequently as- sociated with clinical and subclinical bovine intra-mammary infection. The virulence factors of staphylococcus have not been widely studied in Algeria. The objective of this study was to determine the frequency of slime production, VanA gene and antiseptic resistance genes in staphylococci strains isolated from bovine mas- titis in Algeria. The study examined 35 Staphy- lococci strains obtained from the inflammatory secretion of mammary glands of cows with mastitis. Slime production was determined by detecting the icaA and icaD genes using the polymerase chain reaction (PCR) method and Congo red agar (CRA) method. The presence of qacAB and qac C antiseptic resistance genes and the VanA resistance gene in these isolates was investigated by PCR. The results of the current study revealed that of the 35 Staphylo- cocci isolates, 42.85% (15/35) and 17.14% (6/35) of the isolates harboured the slime production gene by analysing icaA and icaD genes, respec- tively and 71.42% (25/35) by the CRA method. However, VanA and antiseptic resistance genes (qacAB and qac C) were not detected in any of the isolates. Therefore, the majority of Staphylo- coccus strains were capable of producing slime, and the CRA detection rate was higher than the PCR method for the biofilm-producing capac- ity of Staphylococcus strains. Thus, the presence of the ica genes in Staphylococcus strains con- firms its role as a virulence factor in the patho- genesis of bovine mastitis.

scholarly journals Competitiveness of Quantitative Polymerase Chain Reaction (qPCR) and Droplet Digital Polymerase Chain Reaction (ddPCR) Technologies, with a Particular Focus on Detection of Antibiotic Resistance Genes (ARGs)

2021 ◽  
Vol 1 (3) ◽  
pp. 426-444
Author(s):  
Sol Park ◽  
Anita Rana ◽  
Way Sung ◽  
Mariya Munir
Keyword(s):  
Polymerase Chain Reaction ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Antibiotic Resistance Genes ◽  
Chain Reaction ◽  
Viral Infectious Disease ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

With fast-growing polymerase chain reaction (PCR) technologies and various application methods, the technique has benefited science and medical fields. While having strengths and limitations on each technology, there are not many studies comparing the efficiency and specificity of PCR technologies. The objective of this review is to summarize a large amount of scattered information on PCR technologies focused on the two majorly used technologies: qPCR (quantitative polymerase chain reaction) and ddPCR (droplet-digital polymerase chain reaction). Here we analyze and compare the two methods for (1) efficiency, (2) range of detection and limitations under different disciplines and gene targets, (3) optimization, and (4) status on antibiotic resistance genes (ARGs) analysis. It has been identified that the range of detection and quantification limit varies depending on the PCR method and the type of sample. Careful optimization of target gene analysis is essential for building robust analysis for both qPCR and ddPCR. In our era where mutation of genes may lead to a pandemic of viral infectious disease or antibiotic resistance-induced health threats, this study hopes to set guidelines for meticulous detection, quantification, and analysis to help future prevention and protection of global health, the economy, and ecosystems.

scholarly journals Diversity ofLeptospiraspp. in Rats and Environment from Urban Areas of Sarawak, Malaysia

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Chai Fung Pui ◽  
Lesley Maurice Bilung ◽  
Kasing Apun ◽  
Lela Su’ut
Keyword(s):  
Polymerase Chain Reaction ◽  
Water Samples ◽  
Urban Areas ◽  
Environmental Samples ◽  
Soil Samples ◽  
Chain Reaction ◽  
Prevalence Studies ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Soil And Water

Various prevalence studies onLeptospirain animals and humans, as well as environmental samples, had been conducted worldwide, including Malaysia. However, limited studies have been documented on the presence of pathogenic, intermediate, and saprophyticLeptospirain selected animals and environments. This study was therefore conducted to detectLeptospiraspp. in rats, soil, and water from urban areas of Sarawak using the polymerase chain reaction (PCR) method. A total of 107 rats, 292 soil samples, and 324 water samples were collected from April 2014 to February 2015. PathogenicLeptospirawas present in 5.6% (6/107) of rats, 11.6% (34/292) of soil samples, and 1.9% (6/324) of water samples. IntermediateLeptospirawas present in 2.7% (8/292) of soil samples and 1.9% (6/324) of water samples. SaprophyticLeptospirawas present in 10.3% (11/107) of rats, 1.4% (4/292) of soil samples, and 0.3% (1/324) of water samples. From this study, 76Leptospiraspp. were isolated. Based on DNA sequencing, the dominantLeptospiraspp. circulating in urban areas of Sarawak are pathogenicLeptospira noguchii, intermediateLeptospira wolffiiserovar Khorat, and saprophyticLeptospira meyeri, respectively. Overall, this study provided important surveillance data on the prevalence ofLeptospiraspp. from rats and the environment, with dominant local serovars in urban areas of Sarawak.

Quantitative Real-Time Polymerase Chain Reaction Detection of BK Virus Using Labeled Primers

2010 ◽  
Vol 134 (3) ◽  
pp. 444-448 ◽  
Author(s):  
Zhengming Gu ◽  
Jianmin Pan ◽  
Matthew J. Bankowski ◽  
Randall T. Hayden
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Polymerase Chain Reaction ◽  
Bk Virus ◽  
Clinical Samples ◽  
Quantitative Detection ◽  
Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain

Abstract Context.—BK virus infections among immunocompromised patients are associated with disease of the kidney or urinary bladder. High viral loads, determined by quantitative polymerase chain reaction (PCR), have been correlated with clinical disease. Objective.—To develop and evaluate a novel method for real-time PCR detection and quantification of BK virus using labeled primers. Design.—Patient specimens (n = 54) included 17 plasma, 12 whole blood, and 25 urine samples. DNA was extracted using the MagNA Pure LC Total Nucleic Acid Isolation Kit (Roche Applied Science, Indianapolis, Indiana); sample eluate was PCR-amplified using the labeled primer PCR method. Results were compared with those of a user-developed quantitative real-time PCR method (fluorescence resonance energy transfer probe hybridization). Results.—Labeled primer PCR detected less than 10 copies per reaction and showed quantitative linearity from 101 to 107 copies per reaction. Analytical specificity of labeled primer PCR was 100%. With clinical samples, labeled primer PCR demonstrated a trend toward improved sensitivity compared with the reference method. Quantitative assay comparison showed an R2 value of 0.96 between the 2 assays. Conclusions.—Real-time PCR using labeled primers is highly sensitive and specific for the quantitative detection of BK virus from a variety of clinical specimens. These data demonstrate the applicability of labeled primer PCR for quantitative viral detection and offer a simplified method that removes the need for separate oligonucleotide probes.

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