Development of a PCR Kit for Detection of <i>Coxiella burnetii</i> in Ukraine

Марущак, Л В; Дерябин, О. Н.; Dedok, L.; Volosyanko, E. V.; Garcavenko, Tatyana

doi:10.1089/vbz.2019.2518

Cited by 4 publications

(4 citation statements)

References 6 publications

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“…Astobiza et al (2012) detected that serological results should be further analyzed with caution, and that other complementary analysis needed to be done (32). Therefore, our results were in expectance since PCR was more sensitive and greatly specific than ELISA in detecting of C. burnetii (33,34). There was a high possibility of false positive or negative ELISA can be resulted due to insufficient blocking of immobilized antigen, antibody instability, seroconversion, and crossreaction of the secondary antibody (35).…”

Section: Discussionmentioning

confidence: 78%

Serological and Molecular Phylogenetic Detection of Coxiella burnetii in Lactating Cows, Iraq

Gharban

Yousif

2020

Iraqi J. Vet. Med.

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This study is carried out to investigate the prevalence of Coxiella burnetii (C. burnetii) infections in cattle using an enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) assay targeting IS1111A transposase gene. A total of 130 lactating cows were randomly selected from different areas in Wasit province, Iraq and subjected to blood and milk sampling during the period extended between November 2018 and May 2019. ELISA and PCR tests revealed that 16.15% and 10% of the animals studied were respectively positive. Significant correlations (P<0.05) were detected between the positive results and clinical data. Two positive PCR products were analyzed phylogenetically, named as C. burnetii IQ-No.5 and C. burnetii IQ-No.6; and then recorded in the National Center for Biotechnology Information (NCBI) under an accession numbers of MN473204.1 and MN473205.1. Comparative identity of the local strains with NCBI-BLAST strains/isolates revealed 97% similarity and 0.1-0.6% of total genetic mutations/changes. NCBI-BLAST Homology Sequence reported high significant identity (P<0.05) between the local, C. burnetii IQ-No.5 and C. burnetii IQ-No.6; strains and C. burnetii 3345937 (CP014354.1) Netherlands isolate at 99.10% and 99.06%, respectively. The current study concluded that the percentage of infected cows with coxiellosis is relatively high, and Coxiella should be listed as abortive pathogen. Therefore, additional studies should be performed including different animals, samples, and regions

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Section: Discussionmentioning

confidence: 78%

Serological and Molecular Phylogenetic Detection of Coxiella burnetii in Lactating Cows, Iraq

Gharban

Yousif

2020

Iraqi J. Vet. Med.

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“…In addition, there is a potential risk of infection in humans through a variety of ways. Individuals involved in animal farming are particularly at risk (20,24). The results of this study indicated the prevalence of C. burnetii in cow and buffalo milk samples in the Urmia region.…”

Section: Discussionmentioning

confidence: 58%

“…It is a sensitive, safe, and specific procedure to detect C. burnetii in various specimens (19). Various target genes were used (20) for specific C. burnetii identification, such as com1 encoding a 27 kDa outer membrane protein, the superoxide dismutase (Sod B) gene, and the heat shock operon that encodes 2 heat shock proteins (htpA and htpB). The other target genes include the macrophage infectivity potentiator protein (cbmip), isocitrate dehydrogenase (icd), and a transposon-like repetitive region of the C. burnetii genome.…”

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confidence: 99%

Molecular Detection of Coxiella burnetii by Nested PCR Method in Cattle and Buffalo Raw Milk, Urmia Region, Iran

Enferadi,

Ownagh,

Mardani

2023

Jentashapir J Cell Mol Biol

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Background: Coxiella burnetii, a pleomorphic coccobacillus with a Gram-negative cell wall and the cause of query (Q) fever. Amongst animals, farm animals, goats, and sheep are the main reservoirs of Q fever. Methods: This study was conducted to outline the presence of C. burnetii in raw milk received from farm animals and buffalo in all 12 months of 2020 within the Urmia region, northwest Iran. A total of 600 milk samples were received from 3 regions by registering the animals' ages. DNA extraction from milk samples was performed. Results: The nested-polymerase chain reaction (PCR) was efficient in the detection of C. burnetii based on the transposable com1 gene. The results showed that 12.33% (95% CI: 9.9 - 15%) of the total samples (12.66% buffalo and 12% in cattle raw milk) were positive for C. burnetii DNA. The prevalence of C. burnetii in raw milk samples was considerably higher in summer (12.66%, P < 0.05, 95% CI: 9.3 - 17%). In addition, the superiority of C. burnetii in livestock milk drastically varied (P < 0.05) amongst age groups. However, it was not significant in buffalo milk samples. Conclusions: The farm animals and buffalo population in Urmia may be taken into consideration as an important parameter in the epidemiology of Q fever.

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“…Polymerase chain reaction (PCR) is one of the impor-tant detection methods to detect various pathogens like SARS CoV-2. Influenza virus, Hepatitis C virus, Coxiella burnetii and many other pathogens [1][2][3][4][5][6][7][8][9]. Moreover there are large number of RNA viruses in ticks, which are needed to be addressed [10].…”

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confidence: 99%

A simple, effective and inexpensive method to isolate the nucleic acid (DNA/RNA) from a single tick for molecular detection of various pathogens

BHATİA,

BAERSCH

2024

The European Research Journal

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Objectives: Ticks are vectors of a number of pathogens, which cause diseases with fatal consequences, therefore it is essential to detect such pathogens from single tick. Molecular methods like polymerase chain reaction (PCR) are offering such possibilities. At present, cumbersome methods involving liquid nitrogen, cutting ticks with scalpels as well as pooled ticks are being used worldwide. Our goal is to develop a reliable and fast method to obtain nucleic acid (DNA/RNA) from the single tick shipped at room temperature to detect various pathogens. Methods: We developed a mechanical crushing method with mini column nucleic acid isolation from a single tick shipped at room temperature with postal or currier service in a letter. PCR detection was done for Borrelia burgdorferi and tick-borne encephalitis virus as examples. Results: This method was used successfully for the isolation of nucleic acid from single tick and later used for PCR detection of B. burgdorferi and tick-borne encephalitis virus on 17 single tick samples as examples, but for last 18 years, this method was used on more than 250 ticks from Germany. Spectrometric values indicate the presence of sufficient yield of DNA and RNA (up to 900 µg/mL per tick) during the isolation. Conclusions: This may be the first report about a number of one single tick cases, which were sent at room temperature in letters with postal services for isolation of the nucleic acid with mini column kit and used later on for PCR detection of various pathogens. This inexpensive and simple method may be used in any laboratory worldwide for monitoring the presence of tick-borne pathogens.

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Development of a PCR Kit for Detection of Coxiella burnetii in Ukraine

Cited by 4 publications

References 6 publications

Serological and Molecular Phylogenetic Detection of Coxiella burnetii in Lactating Cows, Iraq

Serological and Molecular Phylogenetic Detection of Coxiella burnetii in Lactating Cows, Iraq

Molecular Detection of Coxiella burnetii by Nested PCR Method in Cattle and Buffalo Raw Milk, Urmia Region, Iran

A simple, effective and inexpensive method to isolate the nucleic acid (DNA/RNA) from a single tick for molecular detection of various pathogens

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