wileyonlinelibrary.com/journal/ijlh Int J Lab Hem. 2018;40:196–200.
© 2017 John Wiley & Sons Ltd
Received: 23 June 2017
Accepted: 13 October 2017
Evaluation of microtitre plate- based Haemoglobin estimation
A. P. Shah | P. T. Patel | B. P. Patel | K. K. Mishra | K. Ghosh
Surat Raktadan Kendra & Research Centre,
(NABH accredited Regional Blood Transfusion
Centre), Surat, Gujarat, India
Prof (Dr). Kanjaksha Ghosh, Surat Raktadan
Kendra & Research Centre, Surat, Gujarat,
Introduction: Haemoglobin estimation is one of the most important clinical investiga-
tions. Many techniques are available to measure haemoglobin; still there is a need for
a haemoglobin assay technique which is cheap, robust and simple and can be used in
field conditions very quickly using figure prick sample. We evaluated a
cyanmethaemoglobin- based haemoglobin estimation using a microtitre plates for the
Methods: Microtitre plate- based haemoglobin estimation was developed using
cyanmethaemoglobin- based assay and was compared with standard haematology
analyser- based haemoglobin estimation in a large number of samples from a popula-
tion of voluntary blood donors. Various tests were performed to evaluate the stability
of colour, variation of the results during duplicate assay on the same days and on dif-
ferent days as well as linearity of the test was performed against broad range of hae-
moglobin values for the new microtitre plate- based technique. Standard statistical test
of significance was applied to validate the assay.
Results: Total 200 samples from in- house and field conditions were evaluated. 10 μL
blood sample in 300 μL Drabkin’s solution provided optimum and comparable results
after 10 minutes of incubation. The colour was stable up to 6 hours, the coefficient of
variation was less than 3%, and the cost per test including everything was less than 3
cent/2P. Turnaround time for 90 samples was only 30 minutes.
Conclusion: Cyanmethaemoglobin- based assay in microtitre plate is feasible, robust,
rapid, cheap and cost- effective method for haemoglobin estimation in field
anemia, cheap method, Drabkin’s reagent, epidemiological study, Haemoglobin, microtiter plate
1 | INTRODUCTION
Anaemia is a public health challenge in India, and it mostly affects women
of reproductive age and preschool children. The District Level Health
Survey (DLHS) 2002- 2004 reports an extremely high burden of anae-
mia with 98% of adolescent girls and around 96% of pregnant women
suffering from this condition.
Estimates of the prevalence of anaemia
depend on the methods used for assessing haemoglobin concentration
and on the cut- off point applied as follows: the cut- off point is different
for different population screened for anaemia.
ment is a reliable indicator for anaemia screening. However, there are
multiple techniques with different applications available for haemoglo-
bin estimation and each of the tests differs in cost, technical expertise
required for the test and robustness of the value obtained.
This study was carried out to develop a cyanmethaemoglobin-
based assay in microtitre plate for estimation of haemoglobin in field
condition where large number of persons/population can be tested
cosy effectively and accurately.