Determining the Need for Repeat Testing of Blood Ethanol Concentration: Evaluation of the Synchron Blood Ethyl Alcohol Assay Kit.
In clinical laboratories, a common practice used to verify tests prior to reporting is repeat testing. Our objective was to evaluate the differences between the results of blood ethanol concentration (BEC) test repetitions and report on the role of repeat testing to prevent reporting of incorrect results.We conducted a retrospective study of data retrieved from the Bursa Yuksek Ihtisas Training and Research Hospital’s document management system by calculating the percentage change between repeated BEC test runs.
To assess for clinical relevance, the bias between two results from the same sample was compared using the 1988 Clinical Laboratory Improvement Amendments’ (CLIA) proficiency testing allowable total error (TEa) limits.From a total of 1,627 BEC tests performed between January 2017 and January 2018, 70% (1,133) were repeat tested. Of these, 830 resulted in BECs between 0-5 mmol/L, of which 237 (28.5%) were above the 25% acceptable TEa.
Two hundred seventy-six BEC test results were greater than >14 mmol/L, and there was a good consensus between the initial and repeat test results (99%). In this group, the mean bias was 0.0% (95%, CI = -9.8-9.8%). However, three of the repeat test results were considered significantly different. There were two discordant results in the 5-14 mmol/L ethanol level, and the mean bias was 2.1% (95%, CI = -15.0-19.1%).The majority of the repeated BEC test values were the same as the baseline value; therefore, there may be limited benefit in continuing such frequent repeated analyses.
Lab-on-nanopaper: An optical sensing bioplatform based on curcumin embedded in bacterial nanocellulose as an albumin assay kit.
Herein, we introduce a nanopaper-based analytical device (NAD) or “lab-on-nanopaper” device for visual sensing of human serum albumin (HSA) in human blood serums, which relies on embedding of curcumin within transparent bacterial cellulose (BC) nanopaper. BC nanopaper is an appropriate candidate to be an excellent platform for the development of optical (bio)sensors due to having exceptional properties such as optical transparency, high flexibility, porosity, biodegradability, and printability. The hydrophilic test zones were created on the fabricated bioplatform through creating the hydrophobic walls via laser printing technology.
The color changes of curcumin embedded in BC nanopaper (CEBC) due to the inhibitory effect of HSA on the curcumin degradation in alkaline solutions, which can be monitored visually (naked eye/Smartphone camera) or spectroscopically using a spectrophotometer, were linearly proportional to the HSA concentration in the range of 10-300 μM and 25-400 μM, respectively. The developed NAD/CEBC as a novel albumin assay kit was successfully utilized to the determination of HSA in human blood serum samples with satisfactory results.
Building upon the fascinating features of BC nanopaper as a very promising bioplatform in optical (bio)sensing applications we are confident “lab-on-nanopaper” devices/NADs, which take the advantages of the nanopaper and also meet the ASSURED criteria, could be considered as a new generation of optical (bio)sensing platforms that are currently based on paper, glass or plastic substrates.
Evaluation of the Serologic Cross-Reactivity between Transmissible Gastroenteritis Coronavirus and Porcine Respiratory Coronavirus Using Commercial Blocking Enzyme-Linked Immunosorbent Assay Kits.
This study compared the performances of three commercial transmissible gastroenteritis virus/porcine respiratory coronavirus (TGEV/PRCV) blocking enzyme-linked immunosorbent assays (ELISAs) using serum samples (n = 528) collected over a 49-day observation period from pigs inoculated with TGEV strain Purdue (n = 12), TGEV strain Miller (n = 12), PRCV (n = 12), or with virus-free culture medium (n = 12).
ELISA results were evaluated both with “suspect” results interpreted as positive and then as negative. All commercial kits showed excellent diagnostic specificity (99 to 100%) when testing samples from pigs inoculated with virus-free culture medium. However, analyses revealed differences between the kits in diagnostic sensitivity (percent TGEV- or PRCV-seropositive pigs), and all kits showed significant (P < IMPORTANCE Current measures to prevent TGEV from entering a naive herd include quarantine and testing for TGEV-seronegative animals.
However, TGEV serology is complicated due to the cross-reactivity with PRCV, which circulates subclinically in most swine herds worldwide. Conventional serological tests cannot distinguish between TGEV and PRCV antibodies; however, blocking ELISAs using antigen containing a large deletion in the amino terminus of the PRCV S protein permit differentiation of PRCV and TGEV antibodies. Several commercial TGEV/PRCV blocking ELISAs are available, but performance comparisons have not been reported in recent research. This study demonstrates that the serologic cross-reactivity between TGEV and PRCV affects the accuracy of commercial blocking ELISAs. Individual test results must be interpreted with caution, particularly in the event of suspect results. Therefore, commercial TGEV/PRCV blocking ELISAs should only be applied on a herd basis.
Characterisation of Chinook salmon (Oncorhynchus tshawytscha) blood and validation of flow cytometry cell count and viability assay kit.
New Zealand Chinook salmon (Oncorhynchus tshawytscha) industry has great potential for growth and expansion. While production is relatively free of health problems, there is limited literature on haematology, and immunological tools to safeguard against possible future health threats. The current study aim was to characterise New Zealand farmed O. tshawytscha peripheral blood cellular composition, develop a micro-volume method to isolate peripheral blood mononuclear cells (PBMCs) and validate a microcapillary flow cytometry assay kit for PBMC cell count and viability assessment. We used light microscopy to characterise peripheral blood and PBMC cellular composition in combination with a flow cytometer Sysmex XT 2000i Haematology Analyser. ImageJ version 1.52 was used for cell size characterisation of freshly stained blood.
The stability of PBMCs stained with the Muse® Cell Count and Viability Assay Kit and the Trypan blue assay stains were studied at 4 °C and 21 °C for 60 min; while the Muse® Cell Count and Viability Assay Kit was validated against the Trypan blue assay haemocytometer chamber to assess PBMC count and viability. Findings showed that O. tshawytscha smolt yearlings had total blood cell counts in the range of 1.9-2.7 × 106 μL-1.
Differential cell counts revealed five cell types, comprising 97.18% erythrocytes, 2.03% lymphocytes, 0.67% thrombocytes, 0.09% monocytes, and unquantifiable neutrophils. Using micro-volumes of blood and Lymphoprep™, we successfully isolated fish PBMCs. Significantly, stained PBMCs remained stable for up to 45 min at 4 °C and 21 °C; while validation of the Muse® protocol showed that this microfluidic instrument delivered more accurate and precise viability results than the haemocytometer. The Muse® protocol is rapid, easy to use, has quick calibration steps, and is suitable for field use to facilitate onsite sample processing. These findings pave the way for future assessments of fish health and in vitro immunological studies in O.