scholarly journals Quality of bulk tank milk samples from Danish dairy herds based on real-time polymerase chain reaction identification of mastitis pathogens

2012 ◽  
Vol 95 (10) ◽  
pp. 5702-5708 ◽  
Author(s):  
J. Katholm ◽  
T.W. Bennedsgaard ◽  
M.T. Koskinen ◽  
E. Rattenborg
Keyword(s):  
Polymerase Chain Reaction ◽  
Dairy Herds ◽  
Chain Reaction ◽  
Milk Samples ◽  
Polymerase Chain ◽  
Bulk Tank ◽  
Polymerase Chain Reaction Identification ◽  
Mastitis Pathogens ◽  
Danish Dairy

scholarly journals Identification of subclinical mastitis caused by Mycoplasma spp. from screenings of bulk tanks

2018 ◽  
Vol 70 (6) ◽  
pp. 1793-1797
Author(s):  
S.F. Joaquim ◽  
F.F. Guimarães ◽  
A. Salina ◽  
N.B. Junqueira ◽  
E.N. Gomes ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Subclinical Mastitis ◽  
Mammary Glands ◽  
Dairy Herds ◽  
Mycoplasma Bovis ◽  
Chain Reaction ◽  
Milk Samples ◽  
Polymerase Chain ◽  
Bulk Tank ◽  
Mycoplasma Spp

ABSTRACT Mastitis caused by Mycoplasma spp., regardless of species, are considered highly contagious pathogens and, usually was not responsive to antimicrobial therapy. Five dairy herds, comprising 489 animals and 1,956 mammary glands, were used in this study. Milk samples were obtained from bulk tanks and subjected to polymerase chain reaction (PCR) for the identification of Mollicutes, Mycoplasma spp., and Mycoplasma bovis. Moreover, individual samples from cases of clinical and subclinical mastitis in quarters of the dairy herds’ animals that yielded a positive PCR upon bulk tank analysis were subjected to molecular analysis. Only one bulk tank was positive for class Mollicutes by PCR. All positive samples classified as mastitis teats had their DNA extracted and tested by PCR for both class Mollicutes and M. bovis. Of these, two (2.08%) were positive for Mycoplasma genus, although none was positive for M. bovis. This result suggests that the PCR of bulk tanks is a viable tool in monitoring and preventing mastitis infections caused by Mycoplasma spp.

Detection of clinical bovine mastitis caused by Mycoplasma bovis in Brazil

2020 ◽  
Vol 87 (3) ◽  
pp. 306-308
Author(s):  
Nathália Brancato Junqueira ◽  
Anelise Salina ◽  
Gabriela Capriogli Oliveira ◽  
Elena Mettifogo ◽  
Sâmea Fernandes Joaquim ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Bovine Mastitis ◽  
Dairy Farms ◽  
Clinical Mastitis ◽  
Dairy Herds ◽  
Mycoplasma Bovis ◽  
Chain Reaction ◽  
Research Communication ◽  
Milk Samples ◽  
Polymerase Chain

AbstractThe work reported in this research communication investigated the occurrence of Mycoplasma bovis (M. bovis) in milk samples from cows with clinical mastitis on dairy farms from seven Brazilian states. We hypothesized that M. bovis was present in bovine clinical mastitis milk in Brazil. A total of 561 milk samples were cultured on Hayflick agar and incubated in a microaerophilic atmosphere at 5% CO2. Polymerase chain reaction (PCR) was performed for the detection of Mycoplasma spp. and Mycoplasma bovis in milk samples. Mycoplasma spp. were isolated in 2% of the milk samples, and Mycoplasma bovis was verified in 3% of the milk samples by PCR. The results showed that Mycoplasma bovis is involved in clinical mastitis in Brazilian dairy herds. We emphasize the need for further studies to investigate the infection by this agent in clinical mastitis cases, particularly in Brazil, due to the lack of knowledge about its prevalence.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

Polymerase Chain Reaction for the Direct Detection of Brucella spp. in Milk and Cheese

2001 ◽  
Vol 64 (2) ◽  
pp. 164-167 ◽  
Author(s):  
G. TANTILLO ◽  
A. DI PINTO ◽  
A. VERGARA ◽  
C. BUONAVOGLIA
Keyword(s):  
Polymerase Chain Reaction ◽  
Direct Detection ◽  
Polymerase Chain Reaction Test ◽  
Chain Reaction ◽  
Sheep Milk ◽  
Reaction Test ◽  
Polymerase Chain ◽  
Quality Of Milk ◽  
Chain Reaction Test

A polymerase chain reaction test was developed to detect Brucella spp. directly in milk and cheese and optimized using primers for the BSCP-31 gene. A total of 46 cheese samples produced with sheep and goats milk were assayed, and Brucella spp. was detected in 46% of them, especially in cheese made from sheep milk. This method is of remarkable epidemiologic interest because it is an indirect test indicating the sanitary quality of milk used in dairy industries. The method showed good sensitivity and specificity. It is faster and less expensive than the conventional bacteriological assays.

scholarly journals PCR - based diagnosis to evaluate the performance of malaria reference centers

2004 ◽  
Vol 46 (4) ◽  
pp. 183-187 ◽  
Author(s):  
Silvia Maria Di Santi ◽  
Karin Kirchgatter ◽  
Karen Cristina Sant'Anna Brunialti ◽  
Alessandra Mota Oliveira ◽  
Sergio Roberto Santos Ferreira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gold Standard ◽  
Blood Smear ◽  
Plasmodium Species ◽  
Thick Blood Smear ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Positive Results ◽  
Ssrrna Gene

Although the Giemsa-stained thick blood smear (GTS) remains the gold standard for the diagnosis of malaria, molecular methods are more sensitive and specific to detect parasites and can be used at reference centers to evaluate the performance of microscopy. The description of the Plasmodium falciparum, P. vivax, P. malariae and P. ovale ssrRNA gene sequences allowed the development of a polymerase chain reaction (PCR) that had been used to differentiate the four species. The objective of this study was to determine Plasmodium species through PCR in 190 positive smears from patients in order to verify the quality of diagnosis at SUCEN's Malaria Laboratory. Considering only the 131 positive results in both techniques, GTS detected 4.6% of mixed and 3.1% of P. malariae infections whereas PCR identified 19.1% and 13.8%, respectively.

scholarly journals Genotyping of Listeria by Polymerase Chain Reaction (PCR) and its Epidemiological Significance

2021 ◽  
Vol 6 (2) ◽  
pp. 115-124
Author(s):  
V. Zyuzin ◽  
◽  
O. Tuzova ◽  
U. Frenkel ◽  
Muntian L. ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Internal Control ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Virulence Markers ◽  
Chip Technology ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Epidemiological Significance

The purpose of the study. The article covers the issues of genotyping of listeria by polymerase chain reaction (PCR) and its epidemiological significance. It is known that molecular genetic methods allow to detect specific microbial pathogens, virulence markers, antimicrobial resistance genes faster and with greater sensitivity than traditional culture methods. Therefore, the development of detection methods and genotyping by polymerase chain reaction (PCR) is relevant. Materials and methods. For the detection and genotyping of Listeria, the technology of DNA chips is becoming increasingly important, which can significantly expand the possibilities of molecular detection. Chip technology can be used to simultaneously identify a whole range of pathogenic microorganisms, to determine genetic virulence markers, the relationship to antibiotics, subtyping, as well as to determine the quality of microorganisms in samples. A simplified version of DNA chip technology is multiplex (numerical) PCR, which is used to detect and genotype listeria. Studies have shown that to detect Listeria spp. using a polymerase chain reaction, it is advisable to use the gene iap (invasive associated protein), known for 6 species of listeria, which encodes a protein P 60 that is common to all species of listeria, including L.murrayi. Computer analysis revealed areas with 100% homology, from which primers were selected for PCR detection of all types of listeria. Areas of genomes characterized by 100% homology were selected for further analysis and labeling of primer sets. The sequences of the constructed primers List 1 and List 2 allowed to identify 6 species of Listeria (L. monocytogenes, L. innocua, L. ivanovii, L. grayi, L. seeligeri, L. welshimeri). Increasing the length of the primer leads to the increasing of specificity of PCR analysis. The greater the length of the primer, the smaller the specific gravity of one error of the unpaired nucleotide. The degree of primer homology is a key parameter that indicates the "quality" of a set of primers. Results and discussion. It is established that a significant disadvantage of the vast majority diagnosed using PCR test systems is the lack of internal control of amplification. The negative result of PCR analysis may be due to the absence in the clinical material of a fragment of the Listeria genome, and the fact that the PCR product was not synthesized for other reasons. They may be as the following ones: operator errors, erroneously determined reaction mixture concentrations and PCR temperature parameters. False-negative results can also be caused by factors that inhibit thermostable DNA polymerase. In its turn, such inhibition of the enzyme responsible for amplification is caused by a very large amount of DNA - template, pre-treatment of clinical samples. It has been shown that 80% of clinical specimens contain a substance that inhibits DNA polymerase. Therefore, it is necessary to use internal control, the positive result of the reaction of which indicates the successful amplification, that is the absence of false positive results. Conclusion. There are several reasons why the accuracy of PCR analysis does not reach 100%. Accuracy depends on the technology (variety) of PCR - the method used (ordinary or fluorescent), detection of amplicons, PCR homogeneous or nested, nested in one test tube or in two test tubes, as well as the level of quality of the survey (primarily on the technical parameters of the amplifier). The test systems used can be used for PCR detection and are recommended as standard primer sets for the detection and cross-species testing of listeria, which is important for the timely implementation of appropriate anti-epidemic measures in listeriosis

scholarly journals Streptococcus agalactiae mastitis of bovine detection by Polymerase Chain Reaction (PCR) test in AL-Diwanyia province

2013 ◽  
Vol 12 (2) ◽  
pp. 101
Author(s):  
Gh. K. A. Al-kuzaay ◽  
Q. H. Kshash
Keyword(s):  
Polymerase Chain Reaction ◽  
Streptococcus Agalactiae ◽  
Specific Primer ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Milk Samples ◽  
Bacteriological Testing ◽  
Polymerase Chain ◽  
Pcr Test

This study was conducted for exam 348 milk samples from (clinically mastitic and other healthy cows) in many areas in AL-Diwanyia province by using CMT and bacteriological testing , which appeared that (64.9%) as percentage of mastitis ( clinically 15.9% , subclinically 84.0% ) Streptococcus agalactiae mastitis 13.2% ( 26.6% clinically , 73.3 % subclinicaly) diagnose by PCR assay by using specific primer (16SrRNA). Streptococcus agalactiae (30 isolates) after classical methods applied for streptococcus agalactiae identification (86 isolates).

scholarly journals Evaluating the bacterial culture technique by Polymerase chain reaction for the diagnosis of Brucella abortus in milk of cows suspected with brucellosis.

2016 ◽  
Vol 40 (1) ◽  
pp. 5-8
Author(s):  
Bashar Sadeq Noomy
Keyword(s):  
Polymerase Chain Reaction ◽  
Brucella Abortus ◽  
Bacterial Culture ◽  
Culture Technique ◽  
Test Results ◽  
Chain Reaction ◽  
Culture Techniques ◽  
Milk Samples ◽  
Polymerase Chain ◽  
Pcr Test

      The aim of this study is to determine the sensitivity of bacterial culture technique in the detection of Brucella abortus in milk samples of aborted cows. Sixty samples of milk were collected from aborted cows during a period which did not exceed two months after the abortion. All of them were positive for rose bengal test. Results showed that Brucella abortus was isolated from 7 out of 60 (11.6%) from the milk of aborted cows, while PCR test showed that 32 out of 60 (53.3%) milk sample contained Brucella abortus. The specificity of culture techniques was 10%, but its sensitivity was only 21.8%. Beside the cautions in dealing with live Brucella abortus (as culture), it is also less sensitive than PCR, though it is better to use PCR technique in the diagnosis of brucellosis in aborted cows milk.

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