Automated counting of nucleated cells in equine synovial fluid without and with hyaluronidase pretreatment

Ekmann, Anita; Rigdal, Marie-Louise; Gröndahl, G.

doi:10.1111/j.1939-165x.2009.00203.x

Cited by 19 publications

(13 citation statements)

References 19 publications

(32 reference statements)

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“…Studies on flow-cytometer cell counting in SF samples are of particular relevance to our results on the Luminex platform (Moreno et al, 2000;Sugiuchi et al, 2005;Ekmann et al, 2010). These studies demonstrate that HAse treatment improved the precision by reducing errors due to high viscosity related flow.…”

Section: Discussionmentioning

confidence: 69%

See 1 more Smart Citation

Hyaluronidase treatment of synovial fluid to improve assay precision for biomarker research using multiplex immunoassay platforms

Jayadev

Rout

Price

et al. 2012

Journal of Immunological Methods

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Section: Discussionmentioning

confidence: 69%

“…These properties can make the analysis of SF from OA joints challenging. Consequently, methods to reduce SF viscosity by dilution and/or hyaluronidase (HAse) treatment are routinely used before a variety of analytic techniques (Song et al, 2012;Mateos et al, 2012;Sugiuchi et al, 2005;de Jager et al, 2007;Ekmann et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Hyaluronidase treatment of synovial fluid to improve assay precision for biomarker research using multiplex immunoassay platforms

Jayadev

Rout

Price

et al. 2012

Journal of Immunological Methods

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“…The powdered hyaluronidase did not add appreciable volume to the sample. Specimens were mixed using an automatic mixer for at least 5 min (Ekmann et al, 2010). …”

Section: Methodsmentioning

confidence: 99%

“…The three categories of TNCC were selected for clinical relevance: WRI (<3000/μL) (MacWilliams and Friedrichs, 2003; Ekmann et al, 2010), mild to moderately elevated cell count (3000–20,000/μL) and markedly elevated cell count (>20,000/μL). Four new observers, blinded to slide identity and TNCC values, performed counts once on each of the same 12 slides.…”

Section: Methodsmentioning

confidence: 99%

Relationship between automated total nucleated cell count and enumeration of cells on direct smears of canine synovial fluid

Dusick

Young

Muir

2014

The Veterinary Journal

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Canine osteoarthritis is a common condition seen in veterinary clinical practice and causes considerable morbidity in dogs as they age. Synovial fluid analysis is an important tool for diagnosis and treatment of canine joint disease and obtaining a total nucleated cell count (TNCC) is particularly important. The low volume of fluid obtained during arthrocentesis is often insufficient for obtaining an automated TNCC, thereby limiting sample interpretation. The aim of the present study was to investigate whether estimation of TNCC in canine synovial fluid could be achieved by performing manual cell counts on direct smears of fluid. Fifty eight synovial fluid samples, taken by arthrocentesis from 48 dogs, were included in the study. Direct smears of synovial fluid were prepared, and hyaluronidase added before cell counts were obtained using a commercial laser-based instrument. A protocol was established to count nucleated cells in a specific region of the smear, using a serpentine counting pattern; mean number of nucleated cells/400× field was then calculated. There was a positive correlation between the automated TNCC and mean manual cell count, with more variability at higher TNCC. Regression analysis was performed to estimate TNCC from manual counts. By this method, 78% of the samples were correctly predicted to fall into one of three categories (within the reference interval, mildly to moderately elevated, or markedly elevated) relative to the automated TNCC. Intra-observer and inter-observer agreement was good to excellent. The results of the study suggest that interpretation of canine synovial fluid samples of low volume can be aided by manual cell counting of direct smears.

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“…Evaluation of the sediment smear along with the protein levels and PCV can characterize the disease process. [38][39][40][41][42][43][44][45][46] The PP (total solids) should be used to determine the pathogenesis of the fluid development. Fluids with low protein levels (less than 1.5 g/dL) are considered pure transudates; this implies that water has leaked from vessels as a result of altered fluid forces, including hydrostatic and oncotic pressures.…”

Section: Fluid Evaluationmentioning

confidence: 99%

Overview of Clinical Pathology and the Horse

Lester¹,

Mollat²,

Bryant³

2015

Veterinary Clinics of North America: Equine Practice

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Automated counting of nucleated cells in equine synovial fluid without and with hyaluronidase pretreatment

Abstract: Automated total cell counts obtained on the Medonic analyzer correlate well with manual counts in equine synovial fluid; however, pretreatment with hyaluronidase is required to minimize error flags. Automated differential counts are not accurate for synovial fluid.

Cited by 19 publications

References 19 publications

Hyaluronidase treatment of synovial fluid to improve assay precision for biomarker research using multiplex immunoassay platforms

Hyaluronidase treatment of synovial fluid to improve assay precision for biomarker research using multiplex immunoassay platforms

Relationship between automated total nucleated cell count and enumeration of cells on direct smears of canine synovial fluid

Overview of Clinical Pathology and the Horse

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