
ISSN: 2690-5760
Okorie HM1, Obeagu Emmanuel Ifeanyi1*, Okpoli Henry Chijindu Henry2 and Chukwu Stella Nchekwubedi3
Received: July 18, 2020 Published: August 07, 2020
Corresponding author: Obeagu Emmanuel Ifeanyi, Department of Medical Laboratory Science, Faculty of Health Sciences, Imo State University, Owerri, Nigeria
DOI: 10.32474/JCCM.2020.02.000137
A comparative study of Enzyme linked immunosorbent assay (ELISA) and rapid test screening method on HIV, HBV, HCV and syphilis among voluntary donors was investigated on 350 subjects in Owerri. They were 250 males and 100 females, all ranging from the age of 21-50 years. Transfusion transmissible infections (TTIS) are the major problem associated with blood transfusion. Accurate estimates of risk of TIIS are essential for monitoring the safety of blood supply and evaluating the efficacy of currently employed screening procedures for immunodeficiency virus (HIV 1 and 2) hepatitis B virus (HBV), hepatitis C virus (HVC) and syphilis using both enzyme linked immunosorbent assay (ELISA) and rapid test screening method. For HIV 1 & 2 screening test, the female donors shows significant increase (P<0.05) between the ELISA and Rapid test method while there is no significant increase (P>0.05) between the ELISA and rapid test method for male donors. Hepatitis B virus screening test shows a significant increase (P<0.05) between ELISA and rapid test method for female donors, while there is no significant increase (P>0.05) between the two methods for male donor. Also hepatitis C Virus screening test shows a significant increase (P<0.05) between the two methods for the female donors and shows no significant increase (P>0.05) for male donors. However, there is significant increase (P<0.05) between ELISA and rapid test method for syphilis screening test among female donors while there is no significant increase (P>0.05) for the male donors. This study shows that there is a difference between the two test method, hence indicated that 30(8.57%) infected units of blood would have been transfused due to false negative results with rapid test method. It was found in this study that prevalence of TTIs were high in females than the males.
Keywords:ELISA; Rapid test; HIV; Hepatitis; Syphilis; Blood transfusion
Blood transfusion and component therapies are well established
and essential medical practice. These therapies however are not
without risk and may lead to the transmission of infectious agent
from donor to recipient. The common infectious agent includes
human immuno deficiency virus (HIV 1 and 2), hepatitis B virus
(HBV), hepatitis C virus (HCV) and syphilis (Schreiber et al., 2008).
In pursue of global blood safety, the world health organization
(WHO) recommends that all blood donation should be screened
for evidence of infection prior to the release of blood and blood
products for clinical use WHO, [1]. According to WHO guidelines,
the screening of all blood donations should be mandatory for IV 1 &
11, HBV, HCV and syphilis.
The prevalence of viral infection in blood donations can be
used as a valuable indicator to assess the safety of blood supply and
potential risk of TTIs.
The pre-donation screening of blood donors for TTIs is the
practice by which a prospective donor is tested for the presence of
one or more of the TTI agent by a single rapid or quick method and donation is deferred if the test is reactive for any of the TIIs markers
Bhawani et al. [2].
Universally, the normal procedure is to administer a standard
questioner, measure the concentration of hemoglobin and weight of
the donor. The donor is then bled if found fit based on the selection
criteria and asked to leave after a period of rest. The donor blood
units are then separated into various components and stored. The
blood is then tested using Elisa assay technique. All those reactive
for any of the TTRIs are appropriately discarded including their
respective components. Those non reactive for viral markers are
appropriately labeled and use for transfusion Lequim [3].
Human immune deficiency virus (HIV 1 & 2), HBV, HCV,
and syphilis are the most important agents causing transfusion
transmissible infectious (TIIs) and they constitute large health care
burdens worldwide Kitchen et al. [4].
Transfusion of TIIs during serological negative window period
still poses a threat to blood donor safety. Therefore, strict selection
of blood donors and comprehensive screening of donor’s blood
using standard methods are highly recommended to ensure the
safety of blood for recipient.
This study shows the importance of serological evidence of TTIS
which may be obtained by testing for HIV 1 & 2 Ag-Ab, HBsAg, HCV
antibody and syphilis antibody in serum individual of voluntary
blood donor to reduce the window period and to increase the assay
sensitivity.
New screening test have been developed which simultaneously
detect antigen and antibody WHO, [1].This research is carried out to
ensure that TTIS are prevented by mandatory screening of donated
blood before transfusion using a highly sensitive and specific test
method (ELISA) to ensure the safety of blood for recipients. The
purpose of this study was to compare the specificity and sensitivity
of ELISA and rapid test screening method on HIV HBsAg, HCV and
syphilis among voluntary blood donors in Owerri.
This study was carried out in Owerri, Imo State Nigeria. The
subjects were representative of only those residents in Owerri
town.
Data was analyzed from 350 blood samples collected, 250 were
males and 100 were female within the ages of 21-50 years. They were all voluntary donors in Owerri. Full industry and physical
examination were performed and recorded for all volunteer blood
donors to review their eligibility for donation, using the donor
record form.
Blood samples were screened for the presence of HIV, HBsAg,
HCV and syphilis using Enzyme linked immunosorbent assay
(ELISA) and rapid test screening method. All data were evaluated
using statistical software.
This study was a retrospective, descriptive and comparative
study of ELISA and rapid test screening method for HIV, HBsAg, HCV
and Syphilis among voluntary donor. 450ml of blood were collected
into blood bag and after donation, a small quality of donated
blood (3-5ml) each were put into the pilot bottles (commercially
prepared EDTA). The samples were centrifuged, separated and
stored at - 700C.
For the detection of HIV 1 & 2, the ultra Rapid test strip of
determine and unigold was used to determine the presence of
HIV 1 & 2 in the donor samples. ACON was used to determine the
presence of HBsAg. While Diaspot rapid test strip was used to
determine the presence of HCV and syphilis antibodies.
These are qualitative membrane strips based immunoassay for
the detection of presence of HIV 1 & 2 antigen-antibody, HBsAg,
HCV and syphilis (Trepemonapallidum) antibodies (lgA& M) in
whole blood, serum or plasma.
All data were evaluated with SPSS (Statistical Package for Social
Sciences) version 20.0. Statistical analysis included descriptive
statistics of percentage and chi-square. A P value of <0.05 was
considered significant.
The experimental result (Table 2) shows the HIV status of the sampled population. From the result it can be seen that 5% of the female individuals tested positive to HIV using the ELISA screening technique while 1% tested positive with the Rapid method. However, 2% of the male individuals tested positive to HIV using ELISA screening method while 0.8% of the male individuals tested positive using Rapid method.
Table 1: Results of HIV, HBsAg, HCV and Syphilis using both ELISA and Rapid test methods among voluntary donors in Owerri. -ve Negative , + Ve Positive.
Table 2: Prevalence of HIV according to gender of the donor using ELISA and Rapid test screening methods
Of the 350 samples tested, 10% of the female individuals tested positive to HBV with ELISA screening method while 5% of the female tested positive with rapid screening kit. On the other hand, 3.6% of male tested positive to HBV using ELISA method and 2% of the male individuals tested positive using Rapid method (Table 3).
Table 3: Prevalence of HBV according to gender of the donor using ELISA and Rapid test screening methods
Of the sampled population, 8% of the female individuals tested positive to HCV, and 4% of the male individuals tested positive to HCV with ELISA method while 3% of female and 2.4% of male tested positive to HCV using Rapid test method of blood screening (Table 4).
Table 4: Prevalence of HCV according to gender of the donor using ELISA and Rapid test screening method.
The percentage of female and male individuals that tested positive to syphilis using ELISA method of blood screening are 5% and 2.4% respectively while that of Rapid screening method are 2% of female and 1.6% of male donors (Table 5).
Table 5: Prevalence of SYPHILIS according to gender of the donor using ELISA and Rapid test screening method.
HIV, HBV, HCV and Syphilis are the greatest threats to blood
safety for transfusion recipients and pose a serious public health
problem. HIV units are of major concern due to carrier state and
complications associated with the infections.
On the evaluation of all the samples in the population, 5% and
2% of both female and male donors were found to be reactive to
HIV with ELISA screening method as oppose to 1 % and 0.8% of
female and male individuals with Rapid method. The inferior
performance of Rapid method in comparison to ELISA has also
been reported by Torane et al. [5] where both methods were used
to screen blood donors for HIV infection and the RDT used missed
some samples confirmed reactive by ELISA. This discordance may
possibly be due to low antibody titres especially in recent infections
where the levels may well be below the detection limit of RDTs but
are picked up by the more sensitive enzyme immunoassay and its
spectrophotometric format of result analysis.
In this study, 5% of female samples that initially tested negative
with the rapid method, tested positive with ELISA technique for
HBS. Similarly 1.6% of the male samples that tested negative with
rapid method tested positive with ELISA method for HBS. This
indicates that ELISA test is more sensitive and superior for the
testing of blood donors for HBS.
The finding is consistent with previous report which indicated
that ELISA technique is superior to rapid method in the diagnosis of hepatitis B virus infection among blood donors. Failure of the
rapid method to detect the presence of markers of infectious viral
diseases may be due; inadequate coating of the antigen, nature of
the antigen used and genetic heterogeneity of the virus Torane et
al. [5].
It is observed that the high false negative results with the
rapid method compared to ELISA in this work is in agreement with
previous report Salawu et al. [6] which indicated that there is risk
of donor blood containing HBV being transfused to patients due to
suboptimal testing HBsAg using rapid kits only.
The rapid test method was found to have sensitivity, specificity,
positive-predictive value and a negative predictive value of 3%
and 2.4% for female and male individuals respectively in HCV. The
samples that tested reactive to HCV by ELISA could not be detected
by the rapid method. Failure of the rapid kits to detect HCV reactive
samples may be due to inadequate coating of the antigens, nature of
the antigens used or genetic heterogeneity of the virus.
Most of these rapid assays use recombinant proteins from the
prototype virus alone. The variants of HCV may differ substantially
in nucleotide sequence from one another and show varied
geographical and epidemiological distributions Kaur et al. [7] and
Robert et al. [8].
From the evaluation of the sample in the population, 5% and
2.4% of both female and male donors were reactive to syphilis
infection using ELISA method as against 2% and 1.6% with rapid
method. This could be the fact that ELISA test method is more
sensitive to pick the lowest syphilis antibody titre than that of the
rapid test method Pickening [9]. ELISA test are generally accurate
test. They are considered highly sensitive and specific compared
with rapid test methods. They have added advantage of not needing
radioisotopes [10].
From the screening point of view, 30 (8.57%) infected units
would have been transfused due to false negative result with the
rapid test method.
Bio chemistry
University of Texas Medical Branch, USADepartment of Criminal Justice
Liberty University, USADepartment of Psychiatry
University of Kentucky, USADepartment of Medicine
Gally International Biomedical Research & Consulting LLC, USADepartment of Urbanisation and Agricultural
Montreal university, USAOral & Maxillofacial Pathology
New York University, USAGastroenterology and Hepatology
University of Alabama, UKDepartment of Medicine
Universities of Bradford, UKOncology
Circulogene Theranostics, EnglandRadiation Chemistry
National University of Mexico, USAAnalytical Chemistry
Wentworth Institute of Technology, USAMinimally Invasive Surgery
Mercer University school of Medicine, USAPediatric Dentistry
University of Athens , GreeceThe annual scholar awards from Lupine Publishers honor a selected number Read More...
The annual scholar awards from Lupine Publishers honor a selected number read more...