Two-site immunoenzymometric assay for thyrotropin in dried blood samples on filter paper.

Abstract We describe a sensitive, simple, and rapid two-site immunoenzymometric assay for thyrotropin in dried blood samples on filter paper, for use in screening for neonatal primary hypothyroidism. In this method, two dried-blood spots of 3 mm diameter (equivalent to about 5.4 microliter of blood) are incubated overnight with anti-thyrotropin-beta-D-galactosidase complex in an anti-thyrotropin coated tube. Then the enzyme activity in the washed tube is determined fluorophotometrically. The range of thyrotropin measurable is 10 to 160 milli-int. units/L blood. Values for thyrotropin in dried blood samples determined by this method and those determined by radioimmunoassay correlated highly (r = 0.96).

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Effect of sample collection on α-galactosidase A enzyme activity measurements in dried blood spots on filter paper

Clinica Chimica Acta ◽

10.1016/j.cca.2009.02.008 ◽

2009 ◽

Vol 403 (1-2) ◽

pp. 159-162 ◽

Cited By ~ 21

Author(s):

Petra Olivova ◽

Kristen van der Veen ◽

Emmaline Cullen ◽

Michael Rose ◽

X. Kate Zhang ◽

...

Keyword(s):

Enzyme Activity ◽

Filter Paper ◽

Dried Blood Spots ◽

Sample Collection ◽

Blood Spots ◽

Activity Measurements

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Prediction of haematocrit in dried blood spots from the measurement of haemoglobin using commercially available sodium lauryl sulphate

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1177/0004563217726809 ◽

2017 ◽

Vol 55 (3) ◽

pp. 363-367 ◽

Cited By ~ 11

Author(s):

G Richardson ◽

D Marshall ◽

BG Keevil

Keyword(s):

Filter Paper ◽

Room Temperature ◽

Dried Blood Spots ◽

Microtitre Plate ◽

Blood Samples ◽

Blood Spots ◽

Fast Estimation ◽

Plate Reader ◽

Elisa Plate ◽

Edta Blood

Background When preparing dried blood spots (DBSs), haematocrit (Hct) can affect the ability of the blood to spread through the filter paper, thus resulting in varying quantities of sample being measured when fixed subpunches of the DBSs are taken. It may be important to predict the sample Hct to correct volume differences. Methods Blood (10 µL) was applied to Perkin Elmer 226® paper. The samples ( n = 165) were allowed to dry for 24 h, and the entire blood spots were cut out. Subpunch analysis was also performed on blood spots prepared from 75 µL EDTA blood, taking 6 mm subpunches centrally and peripherally from the spots ( n = 59). The spots were eluted with 100 µL water, and a 10 µL aliquot of lysate was added to sulfolyser reagent (80 µL) in a microtitre plate. Hb was measured at 550 nm using an ELISA plate reader. DBS samples were compared against blood samples measured on a routine Sysmex XN-9000 analyser. Results The Passing and Bablock regression showed Hct (DBS-predicted) = 0.99 Hct (Sysmex) −0.02, R2 = 0.87. Intra-assay imprecision measured at Hct values of 0.27, 0.40 and 0.52, gave CVs of 4.1%, 2.8% and 4.2%, respectively. Inter-assay imprecision showed CVs of 6.2%, 5.2% and 4.2%, respectively. DBS samples were stable for up to two days at 60℃, one month at room temperature and six months at 4℃. Conclusion This method provides a simple and fast estimation of predicted Hct in dried blood spots.

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Gender identification in Chicken (Gallus gallus) by PCR using whole blood and dried blood spot on filter paper as template: without prior DNA isolation

10.1101/046888 ◽

2016 ◽

Author(s):

S. Dhanasekaran ◽

G. Dhinakar Raj ◽

A. R. Vignesh ◽

S. T. Selvan ◽

B. Prakash ◽

...

Keyword(s):

Sex Determination ◽

Dna Extraction ◽

Filter Paper ◽

Whole Blood ◽

Dried Blood Spots ◽

Sex Identification ◽

Gender Identification ◽

Blood Samples ◽

Blood Spots ◽

Blood Spot

AbstractAccurate sex identification of pure line chickens in their early age has significant economic impact in breeding industry. In the recent years, range of Polymerase Chain Reaction (PCR) based sex determination techniques are routinely used to identify the sex of parent lines in breeding industries, however purified DNA is a prerequisite. Hence this study was aimed to develop a rapid and inexpensive PCR based gender identification method for chicken using whole blood samples and dried blood spots as template for PCR without DNA extraction. In addition, practicability of two W-chromosome specific gene targets in chicken for sex determination also characterised. Successful amplification of sex specific fragments and an internal control was achieved with the range of 0.125μl and 0.250μl volume of whole blood on filter paper (~1 mm) prepared from chicken and dried blood spot. This technique does not require DNA extraction, freeze/thawing of blood samples, pre-treatment with any reagents, dilution of whole blood or dried blood spots on filter paper. It can be carried out with commercially available Taq polymerase enzymes with increased concentration of MgCl2 (3 mM) and 0.5% of DMSO without optimisation of PCR buffers. In conclusion, as compared to the existing PCR based sex identification techniques, the present approach is relatively economic, time saving, requires minimal steps and eliminates the need for DNA extraction.

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