Artifactual changes in the haematocrit buffy coat of stored anti-coagulated blood samples of farm animals

2009 ◽  
Vol 19 (5) ◽  
pp. 493-497 ◽  
Author(s):  
John I. Ihedioha ◽  
Patrick E. Aba
Keyword(s):  
Buffy Coat ◽  
Farm Animals ◽  
Blood Samples ◽  
Artifactual Changes

scholarly journals Comparison of capillary, venous and buffy coat blood samples in detecting Plasmodium species among malaria suspected patients attending at Hamusite health center. A cross-sectional study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Getu Abeje ◽  
Woyneshet Gelaye ◽  
Getaneh Alemu
Keyword(s):  
Health Center ◽  
Detection Rate ◽  
Venous Blood ◽  
Cross Sectional Study ◽  
Buffy Coat ◽  
Blood Film ◽  
Capillary Blood ◽  
Sectional Study ◽  
Cross Sectional ◽  
Blood Samples

Abstract Background Both capillary and venous blood samples have been interchangeably used for the diagnosis of malaria in Ethiopia. However, Plasmodium parasites are thought to be more concentrated in capillary than in venous blood. Hence, selecting a sample source where parasites are more concentrated is indispensable approach in order to maximize the accuracy of blood film microscopy. Therefore, the present study aimed to compare the detection rate and the parasitemia level of Plasmodium species from conventional capillary and venous blood films, and buffy coat preparations. Methods A facility based cross-sectional study was conducted from Feburary to March 2020 among 210 febrile patients attending Hamusite health center, northwest Ethiopia. Capillary and venous blood samples were collected and buffy coat was prepared from each sample. Thin and thick blood films were prepared, stained, and examined microscopically following standard protocol. Data were analysed using Statistical Package for Social Sciences Software version 20 and Med-Calc software version 19.3. Results Capillary blood buffy coat (61/210, 29.0%) had significantly higher detection rate as compared to capillary (48/210, 22.9%) and venous (42/210, 20.0%) blood films (p < 0.001). However, no significant difference was observed between capillary and venous blood films (p = 0.070) in detecting Plasmodium species. The highest and the lowest mean asexual stage parasite counts were found in capillary blood buffy coat (4692.88) and venous blood (631.43) films, respectively showing significant variations (p < 0.001). Mean gametocyte count was also highest in capillary blood buffy coat (3958.44). As compared to capillary blood buffy coat, the sensitivity of venous blood buffy coat, capillary blood film and venous blood film were 73.8, 78.7, 68.9%, respectively. Conclusion Capillary blood buffy coat samples showed the highest sensitivity in detecting and quantitating malaria parasites that its use should be promoted in clinical settings. However, conventional capillary and venous blood films could be used interchangeably.

scholarly journals Evaluation of a Preclinical Blood Test for Scrapie in Sheep Using Immunocapillary Electrophoresis

2006 ◽  
Vol 89 (3) ◽  
pp. 720-727 ◽  
Author(s):  
Roy Jackman ◽  
David J Everest ◽  
Mary Jo Schmerr ◽  
Mohammed Khawaja ◽  
Pat Keep ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Prion Protein ◽  
Clinical Signs ◽  
Test System ◽  
Buffy Coat ◽  
Infected Sheep ◽  
Test Method ◽  
Peptide Sequence ◽  
Transmissible Spongiform Encephalopathies ◽  
Blood Samples

Abstract An analytical method is described for detection of endogenous disease-associated prion protein in the buffy coat fraction from the blood of sheep infected with scrapie. The method has been improved and evaluated for its performance in the preclinical diagnosis of ovine transmissible spongiform encephalopathies. The test system uses a protocol for sample preparation that includes extraction and concentration and a test method that uses a liquid-phase competitive immunoassay for prion protein. Antibodies directed to a peptide sequence at the C-terminus of the prion protein (PrP) and a fluorescein-labeled peptide conjugate are used in the assay. Free zone capillary electrophoresis with laser-induced fluorescence for detection is used to separate the antibody-bound fluorescently labeled peptide and free labeled peptide. In this assay, the PrP competes with the fluorescently labeled peptide for limited antibody binding sites, which results in a reduction of the peak representing the immunocomplex of the antibody bound to the fluorescently labeled peptide. When blood samples from scrapie-infected sheep aged 712 months and of the scrapie-susceptible PrP genotypes VRQ/VRQ and VRQ/ARQ were analyzed, the abnormal PrP was found in blood samples. These results correlated with the post-mortem diagnosis of scrapie. The sheep were preclinical and appeared normal at the time of testing but later died with clinical disease approximately 12 months after testing. In older animals, and those with clinical signs, a smaller percentage of animals tested positive. This study has demonstrated that this technology can be used as a sensitive, rapid preclinical test to detect the disease-associated PrP in the blood of scrapie-infected sheep. Improvements in the extraction protocol and capillary electrophoresis conditions will enhance the robustness of this test.

scholarly journals Cord blood buffy coat DNA methylation is comparable to whole cord blood methylation

2017 ◽  
Author(s):  
John Dou ◽  
Rebecca J. Schmidt ◽  
Kelly S. Benke ◽  
Craig Newschaffer ◽  
Irva Hertz-Picciotto ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Blood Cell ◽  
Cord Blood ◽  
Whole Blood ◽  
Cell Types ◽  
Buffy Coat ◽  
Sample Type ◽  
Blood Samples ◽  
Cell Composition ◽  
Cell Counts

AbstractBackgroundCord blood DNA methylation is associated with numerous health outcomes and environmental exposures. Whole cord blood DNA reflects all nucleated blood cell types, while centrifuging whole blood separates red blood cells by generating a white blood cell buffy coat. Both sample types are used in DNA methylation studies. Cell types have unique methylation patterns and processing can impact cell distributions, which may influence comparability.ObjectivesTo evaluate differences in cell composition and DNA methylation between buffy coat and whole cord blood samples.MethodsCord blood DNA methylation was measured with the Infinium EPIC BeadChip (Illumina) in 8 individuals, each contributing buffy coat and whole blood samples. We analyzed principal components (PC) of methylation, performed hierarchical clustering, and computed correlations of mean-centered methylation between pairs. We conducted moderated t-tests on single sites and estimated cell composition.ResultsDNA methylation PCs were associated with individual (PPC1=1.4x10-9; PPC2=2.9x10-5; PPC3=3.8x10-5; PPC4=4.2x10-6; PPC5=9.9x10-13), and not with sample type (PPC1-5>0.7). Samples hierarchically clustered by individual. Pearson correlations of mean-centered methylation between paired individual samples ranged from r=0.66 to r=0.87. No individual site significantly differed between buffy coat and whole cord blood when adjusting for multiple comparisons (5 sites had unadjusted P<10-5). Estimated cell type proportions did not differ by sample type (P=0.86), and estimated cell counts were highly correlated between paired samples (r=0.99).ConclusionsDifferences in methylation and cell composition between buffy coat and whole cord blood are much lower than inter-individual variation, demonstrating that both sample preparation types can be analytically combined and compared.

Artifactual changes in hematological variables in equine blood samples stored at different temperatures and various anticoagulants

2010 ◽  
Author(s):  
Maysam Tehrani-Sharif ◽  
Mehrdad Ameri ◽  
Sogand Moshfeghi ◽  
Hamid Sharifi ◽  
Seyed Mohammad Hoseini ◽  
...  
Keyword(s):  
Blood Samples ◽  
Different Temperatures ◽  
Artifactual Changes

scholarly journals Rapid Diagnosis of Bacteremia in Adults Using Acridine Orange Stained Buffy Coat Smears

1990 ◽  
Vol 1 (1) ◽  
pp. 7-10
Author(s):  
Mark Miller ◽  
Jack Mendelson
Keyword(s):  
Blood Culture ◽  
Acridine Orange ◽  
Buffy Coat ◽  
Blood Cultures ◽  
Rapid Screening ◽  
Blood Culture System ◽  
Blood Samples ◽  
Significant Difference ◽  
Rapid Screening Test ◽  
Low Sensitivity

The use of acridine orange stained buffy coat smears was assessed as a rapid screening test for bacteremia in adults. A total of 356 consecutive blood cultures were submitted with simultaneous anticoagulated blood samples, from which a buffy coat smear was prepared and stained with acridine orange (100 mg/L; pH 3.0). Forty-one of 356 blood samples (12%) yielded organisms in the blood culture system. Compared to blood culture, the overall sensitivity of acridine orange stained buffy coat smears was 16%, specificity 88%, and positive predictive value 13%. There was no statistically significant difference in performance of the test among patients who had fever greater than 39°C and/or shock. The low sensitivity and specificity of the test makes it unsuitable as a means of rapid screening for adults with suspected bacteremia.

scholarly journals Development of a Method to Detect Mycobacterium paratuberculosis in the Blood of Farmed Deer Using Actiphage® Rapid

2021 ◽  
Vol 8 ◽  
Author(s):  
Anton Kubala ◽  
Tania M. Perehinec ◽  
Catherine Evans ◽  
Andrea Pirovano ◽  
Benjamin M. C. Swift ◽  
...  
Keyword(s):  
Gradient Centrifugation ◽  
Elisa Test ◽  
Buffy Coat ◽  
Test Results ◽  
Blood Samples ◽  
Diagnostic Potential ◽  
Ficoll Gradient ◽  
Blood Lysis ◽  
Small Set ◽  
Cattle Blood

Mycobacterium avium subsp paratuberculosis (MAP) is the causative agent of Johne's disease, which is an economically and clinically relevant pathogen for commercial deer production. The purpose of this study was to develop a method that could be used to rapidly detect MAP infection in deer using the Actiphage Rapid blood test. This test has previously been used to detect MAP in cattle blood following the purification of buffy coat using Ficoll gradients, however this method is quite laborious and costly. The purpose of this study was to develop a simpler method of blood preparation that was also compatible with deer blood and the Actiphage test. Initially differential lysis of RBCs using Ammonium Chloride-Potassium (ACK) blood lysis buffer was compared with the Ficoll gradient centrifugation method using cattle blood samples for compatibility with the Actiphage reagents, and it was found that the simpler ACK method did not have an impact on the Actiphage test reagents, producing an equivalent sensitivity for detection of low levels of MAP. When the two methods were compared using clinical blood samples from farmed deer, the ACK lysis method resulted in a cleaner sample. When a blinded test of 132 animals from 4 different production groups was carried out, the majority of the positive test results were found to be from animals in just one group, with a small number identified in a second group. The test results were found to be reproducible when a small set of positive animals were tested again 1 month after their initial testing. Finally a set of negative animals which had been previously screened using an ELISA test, all animals gave a negative Actiphage result. This study shows that this improved sample preparation method and Actiphage blood testing can be used to test blood samples from deer, and the full diagnostic potential of the method can now be evaluated.

scholarly journals Serological and Molecular Identification of Brucella spp. in Pigs from Cairo and Giza Governorates, Egypt

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 248 ◽  
Author(s):  
Khan ◽  
Melzer ◽  
El-Soally ◽  
Elschner ◽  
Mohamed ◽  
...  
Keyword(s):  
Real Time Pcr ◽  
Brucella Melitensis ◽  
Control Strategies ◽  
Farm Animals ◽  
Effective Control ◽  
Serum Samples ◽  
Blood Samples ◽  
Male And Female ◽  
Molecular Assays ◽  
Brucella Suis

Brucellosis is considered as endemic disease of animals and humans since thousands of years in Egypt. However, brucellosis in pigs has never been reported in Egypt. Thus, serological and molecular assays were applied to detect anti-Brucella antibodies and DNA in serum samples collected from pigs. In total 331 blood samples collected from male and female pigs at slaughterhouses of Cairo and Giza governorates were investigated using Brucella c- and i-ELISA and Brucella real-time PCR. Anti-Brucella antibodies were detected in 16 (4.83%) and 36 (10.8%) sera by i-ELISA and c-ELISA, respectively. Brucella DNA was detected in 10 (3.02%) seropositive samples and identified as Brucella melitensis (7/10) and Brucella suis (3/10). A higher prevelance was found in boars. This is the first study investigating pig brucellosis in Egypt. The results of this study will raise awareness for brucellosis in these farm animals and will help to develop effective control strategies.

scholarly journals An outbreak of caprine toxoplasmosis - investigation and case report

2018 ◽  
Vol 48 (5) ◽  
Author(s):  
José Mauricio Ferreira Neto ◽  
Fernanda Pinto Ferreira ◽  
Ana Carolina Miura ◽  
Jonatas Campos de Almeida ◽  
Felippe Danyel Cardoso Martins ◽  
...  
Keyword(s):  
Management Practices ◽  
Clinical Signs ◽  
Antibody Test ◽  
Buffy Coat ◽  
Soil Samples ◽  
Dairy Goat ◽  
Blood Samples ◽  
Lactating Goats ◽  
Polymerase Chain ◽  
Immunofluorescence Antibody Test

ABSTRACT: The present study aimed to investigate an abortion outbreak in a dairy goat herd in the municipality of Arapoti, Parana, Brazil. At the beginning of the outbreak, blood samples were collected from 33 goats with clinical signs; later, of the whole goat herd, two cats and two dogs. Milk samples were collected from 78 lactating goats. Four environmental soil samples and four samples of feed residue from goat feeders were collected too. Immunofluorescence antibody test (IFA) was used for serodiagnosis, the molecular analysis was conducted by means of the polymerase chain reaction (PCR), for the isolation of the etiological agent the bioassay was used. The results of the IFA revealed that 76.53% (137/179) of the goats, two dogs and two cats were seropositive for Toxoplasma gondii. Bioassay revealed one buffy coat and two milk sample having viable T. gondii. In the PCR, 11 whole blood samples, eight milk, three feeder troughs, and all soil samples were positive. The findings of the present study confirmed an outbreak caused by environmental contamination (of soil and feed) with T. gondii oocysts that could have been shed by kittens that lived on the farm and had access to the stock of goat food, facilitating this contamination, which reinforces the need for veterinary assistance and good management practices on farms.

scholarly journals Quantitative Near-Infrared Imaging of Platelets in Platelet-Rich Fibrin (PRF) Matrices: Comparative Analysis of Bio-PRF, Leukocyte-Rich PRF, Advanced-PRF and Concentrated Growth Factors

2020 ◽  
Vol 21 (12) ◽  
pp. 4426 ◽  
Author(s):  
Hachidai Aizawa ◽  
Tetsuhiro Tsujino ◽  
Taisuke Watanabe ◽  
Kazushige Isobe ◽  
Yutaka Kitamura ◽  
...  
Keyword(s):  
Growth Factors ◽  
Near Infrared ◽  
Infrared Imaging ◽  
Buffy Coat ◽  
Gradient Theory ◽  
Blood Samples ◽  
Platelet Rich Fibrin ◽  
Plasma Fraction ◽  
Distal Surface ◽  
Concentrated Growth Factors

Platelet-rich fibrin (PRF) is a fibrin matrix enriched with platelets. The PRF matrix is thought to form a steep gradient of platelet density around the region corresponding to the buffy coat in anticoagulated blood samples. However, this phenomenon has not yet been proven. To visualize platelet distribution in PRF in a non-invasive manner, we utilized near-infrared (NIR) imaging technology. In this study, four types of PRF matrices, bio-PRF, advanced-PRF (A-PRF), leukocyte-rich PRF (L-PRF), and concentrated growth factors (CGF) were compared. Blood samples collected from healthy, non-smoking volunteers were immediately centrifuged using four different protocols in glass tubes. The fixed PRF matrices were sagittally divided into two equal parts, and subjected to modified immunohistochemical examination. After probing with NIR dye-conjugated secondary antibody, the CD41+ platelets were visualized using an NIR imager. In L-PRF and CGF, platelets were distributed mainly on and below the distal surface, while in bio-PRF and A-PRF, platelet distribution was widespread and homogenous. Among three regions of the PRF matrices (upper, middle, and lower), no significant differences were observed. These findings suggest that platelets aggregate on polymerizing fibrin fibers and float up as a PRF matrix into the plasma fraction, amending the current “gradient” theory of platelet distribution.

Preparation of the Buffy Coat From Small-Volume Blood Samples For Ultrastructural Study

1985 ◽  
Vol 43 ◽  
pp. 712-713
Author(s):  
M. J. Mitchell ◽  
E. J. Mirro ◽  
H. E. Black ◽  
E. Schwartz
Keyword(s):  
Peripheral Blood ◽  
Small Volume ◽  
Electron Microscopic Examination ◽  
Buffy Coat ◽  
Electron Microscopic ◽  
Blood Samples ◽  
Toxicological Studies ◽  
Volume Blood ◽  
Toxicological Research ◽  
Early Indication

Electron microscopic examination of peripheral blood is a valuable technique in toxicological research. Blood can be collected for examination several times during a toxicological study. Compound-induced cellular changes detected in peripheral blood cells prior to the terminal necropsy may be an early indication of potential compound-related toxicity.The routine use of small rodents in toxicological studies presents a problem; the amount of blood available at interim collections is minimal. The following technique has been refined for the purpose of examining leukocytes in small blood samples (1 cc or less). Leukocytes are concentrated so that a maximal number per sample can be examined.

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