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*Corresponding author: E-mail: akeler@abuad.edu.ng; 
 
 
 

Asian Journal of Immunology 
 
3(1): 199-207, 2020; Article no.AJI.57099 
 

 
 

 

 

Seroprevalence of Rubella IgG and IgM Antibodies 
among Pregnant Women Attending Antenatal Clinic 

in Federal Teaching Hospital Ido-Ekiti, Nigeria 
 

Richard Yomi Akele1*, Baget Nerat Pam1, Bernard Oluwapelumi Oluboyo1,  
Seyi Samson Enitan2, Janet Funmilayo Akinseye1  

and Funmilayo Ajoke Adewumi3 
 

1
Department of Medical Laboratory Science, College of Medicine and Health Science, Afe Babalola 

University, Ado-Ekiti, Nigeria. 
2
Department of Medical Laboratory Science, Babcock University, Ilisan-Remo, Ogun State, Nigeria. 

3
Department of Microbiology and Parasitology, College of Medicine, Ekiti State University, Ado-Ekiti, 

Nigeria. 
 

Authors’ contributions  
 

This work was carried out in collaboration among all authors. Authors RYA and BNP designed the 
study, wrote the protocol, collected data, performed analysis and wrote the first draft of the 

manuscript. Authors BOO and SSE performed the statistical analysis and restructured the manuscript. 
Authors JFA and FAA managed literature searches. All authors read and approved the final 

manuscript. 
 

Article Information 
 

Editor(s): 
(1) Dr. Darko Nozic, Professor, University of Belgrade, Serbia. 
(2) Dr. Jaffu Othniel Chilongola, Tumaini University, Tanzania. 

Reviewers: 
(1) Victor B. Oti, Nasarawa State University, Nigeria. 

(2) Simeon Achunam Nwabueze , Namdi Azikiwe University, Nigeria. 
Complete Peer review History: http://www.sdiarticle4.com/review-history/57099 

 
 
 
 

Received 22 March 2020  
Accepted 29 May 2020 

Published 04 June 2020 

 
 

ABSTRACT 
 

Aim: Though Rubella is vaccine-preventable and enlisted on the expanded program on 
immunization (EPI) list, vaccination and testing are not routinely practiced in Ido-Ekiti. There is also 
paucity of epidemiological data on the prevalence of rubella infection at Ido-Ekiti, hence the study 
aimed at carrying out a serosurvey to generate epidemiological data for this location. 
Study Design: This was a cross-sectional study. 
Place and Duration of Study: The study was carried out between October 2018 and January 
2019 at the Antenatal Clinic of Federal Teaching Hospital Ido-Ekiti (FETHI), Ekiti State, Nigeria. 

Original Research Article 



 
 
 
 

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Methodology: One hundred and eighty four consenting pregnant women attending antenatal clinic 
at FETHI were enrolled. Structured questionnaire was administered to collect Sociodemographic 
data and sera samples were also collected to determine seroprevalence of rubella IgG and IgM 
antibodies using the enzyme linked immunosorbent assay (ELISA) method. Prevalence rate was 
calculated and chi square value was determined. 
Results: Of the 184 sera samples analyzed 176 (95.7%) and 22 (12%) were seropositive for 
rubella IgG and IgM respectively. Twenty (11%) of those seropositive for IgM were also positive for 
IgG and only 2(1%) were positive only for IgM. Prevalence rate for IgG was highest among 26-
30years (98.3%) and 31-35years (18.8%) for IgM. Age had no-significant effect (p> 0.05) on 
seroprevalence distribution. In like manner, level of formal education, knowledge of the virus, and 
occupation had no significant effect (p> 0.05) on prevalence of the virus. However parity 
significantly (p< 0.05) influenced the pattern of serostatus for both IgG and IgM. 
Conclusion: The high prevalence and similar distribution pattern irrespective of sociodemographic 
features of rubella virus in this study area suggests its endermicity and continuous transmission in 
the area. This emphasizes the need to implement routine immunization of children and susceptible 
women of child bearing age against rubella virus. 

 

 
Keywords: Seroprevalence; rubella IgG and IgM; pregnancy; congenital rubella syndrome; Ido-Ekiti; 

Nigeria. 
 

1. INTRODUCTION 
 
Rubella is a contagious viral infection 
characterized by distinctive macules of 
erythematous rash popularly called German 
measles. It is an enveloped positive-sense 
single-stranded ribonucleic acid (RNA) virus 
belonging to the family Togaviridae, genus 
Rubivirus [1-3]. Rubella is transmitted 
congenitally and by respiratory aerosols, 
transmission occurring mainly in children and 
young adults. The contagious period of the virus 
is approximately 5 to 7-days pre and 3 to 5-days 
post manifestation of clinical symptoms [4]. Upon 
inhalation of virus loaded droplets, the virus 
infects cells of the mucosal membranes of the 
upper respiratory tract and replicates 
consequently spreading to regional lymph nodes. 
[4]. Infection is mainly subclinical, alternatively 
causing a self-limiting illness with low-grade 
fever, lymphadenopathy, and skin rash.  Of major 
concern is the fact that infection in pregnancy 
often result in congenital rubella infection which 
causes miscarriages, stillbirth, abortion, or 
congenital rubella syndrome (CRS) of the infants. 
Congenital rubella syndrome encompasses 
cardiac, cerebral, ophthalmic, and auditory 
defects [5-7]. It is estimated that infection with 
the virus in the first trimester or shortly                   
before pregnancy may result in approximately 
90% of fetuses being infected and 100% of  
these infected fetuses will come down with 
congenital deformities, often resulting in 
miscarriage [4,8-9]. It has been reported that as 
pregnancy progresses, the risk of CRS 
decreases [9]. 

In Nigeria, a high prevalence rate of rubella has 
been reported in some areas. For example, 77% 
in Lagos [10], 97.9% in Zaria [11] and 93.1% in 
Abakaliki [12]. Seroepidermiological studies in 
Ethiopia also has reported a similarly high 
prevalence, 91% in Addis Ababa [13] and 86.3% 
[14] in Southern Ethiopia. 

 
Rubella is a vaccine-preventable viral infection 
[15] considered as an eradicable infection. Based 
on this fact, some vaccination programs 
implemented between 1996 and 2010 in some 
high income, middle income and low-income 
countries have in the past been able to 
considerably reduce global CRS cases [16,17]. 
This has stirred the World Global Assembly to 
endorse the Global Measles and Rubella 
Strategic plan in April 2012 [18] which is targeted 
at eliminating rubella in at least 5 WHO regions 
by 2020 [19]. However, routine administration of 
the rubella vaccine is yet to be implemented in 
many areas in Nigeria most especially in Ido-Ekiti 
were this study was carried out. Routine 
diagnosis for the rubella virus is also not 
practiced in this location just as the case is in 
many other locations in Nigeria. Thus, a paucity 
of empirical data on serosurvey of rubella. 
Though some researchers have conducted 
serosurveillance in other locations, this is the first 
to be carried out in Ido-Ekiti and environs. This 
makes it pertinent to conduct this serosurvey in 
this location as findings may propel informed 
decision making and definition of the best 
interventional approach by those saddled              
with such responsibilities. Findings of such 
investigation may be cardinal to the estimation of 



 
 
 
 

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the incidence of CRS and conversely CRS 
burden at Ido-Ekiti and its environs. This study 
was therefore designed to find out the 
seroprevalence of anti-rubella IgG antibodies (a 
marker of prevalence rate), anti-rubella IgM (a 
marker of incidence rate), and some 
sociodemographic cardinals that may influence 
the distribution of the disease among pregnant 
women in Ido-Ekiti. 
 

2. METHODOLOGY 
 

2.1 Study Area and Population 
 

This cross-sectional study was conducted at the 
antenatal clinic of Federal Teaching Hospital Ido-
Ekiti (FETHI).  FETHI is a referral hospital that 
receives patients from all over Ekiti State and 
neighboring States like Kwara and Kogi State. It 
is sited in a sub-urban community located in Ido-
Osi Local Government Area of Ekiti State 
Nigeria, on latitude 7.843093 and 5.182314 (Find 
Coordinates.https://www.distancesto.com/coordi
nates/ng/ido-ekiti-latitude-longitude/history/759 
44.htmllast accessed 04 April 2020) [20]. 
 

The study population comprised of 184 pregnant 
women at various stages of pregnancy that 
consented to participate in the study.  
 

2.2 Sample Collection and Processing 
 

A brief enlightenment on the basis, benefits and 
demerit of the study was given and then a written 
consent obtained from volunteers. A structured 
questionnaire was administered to those who 
indicated interest in participating in the study. 
The questionnaire was designed to collect 
sociodemographic data and data on possible risk 
factors that may predispose subjects to infection 
[14]. After duly filling the questionnaire, 3 mls of 
venous blood was collected into a labeled plain 
tube using standard aseptic procedures. Blood 
samples were allowed to clot, aseptically, 
dislodged, and centrifuged at 2,500 rpm for 5 
minutes [21]. Sera were then separated into well-
labeled cryovials, transported to the Medical 
Microbiology Laboratory of Afe Babalola 
University, Ado-Ekiti on ice packs. The samples 
were stored at -20°C until ready for laboratory 
analysis. Samples were collected over a period 
of four months.   
 

2.3 Sample Analysis 
 

Sera samples were analyzed for rubella IgG and 
IgM antibodies in the Medical Microbiology 
Laboratory of Afe Babalola University Ado-Ekiti 
using the enzyme-linked immunosorbent assay 

(ELISA) method. Rubella IgG and IgM pre-
coated ELISA kits were obtained from Dia Pro. 
Diagnostic Bioprobes Srl®, Columella Millano, 
ITALY. The analysis was carried out according to 
the manufacturer’s instructions. 
 

Briefly, reagents provided were allowed to attain 
room temperature for 15 minutes before use. The 
40X wash buffer was diluted with distilled water 
using a ratio of 1:40 before use. The micro-titer 
plate template was set up with 1 well as blank, 2 
wells as negative control and 2 wells as the 
positive control. 10 µl of sera sample and 90µl 
sample diluent were dispensed into the 
respective wells except for the blank well, 
negative control well and positive control well. 
100 µl of the negative and positive controls were 
dispensed into their wells respectively. The 
content was mixed by vibrating the plate gently. 
The microplate was covered with a sealing paper 
and incubated in a microplate incubator 
(MARVOTECH PLATE INCUBATOR, CHINA) at 
37°C for 60 minutes [21]. After incubation, the 
microplate was washed five times using wash 
buffer. 100 µl of Horseradish peroxidase enzyme 
(HRP) conjugate was added to each well except 
the blank; the microplate was covered with a 
sealing paper and also incubated in a microplate 
incubator at 37°C for 15 minutes. After 
incubation, the microplate was washed five times 
with the diluted wash buffer in an automatic plate 
washer (MARVOTECH PLATE WASHER, China) 
[21]. 50 µl of substrate solution A and B were 
added to each well respectively and were mixed; 
the plate was covered and incubated at 37°C for 
15 minutes. 50 µl of stop solution was added to 
each well and mixed. The absorbance was read 
in an ELISA reader machine (MARVOTECH 
ELISA READER, China) at a wavelength of 450 
nm [21].  
 

2.31 Interpretation of rubella IgG/IgM result 
 

If the mean negative control O.D ≤ 0.1and the 
mean positive control O.D ≥ 0.8, the test is valid.  
 

Cut-off O.D = the mean O.D value of the 
negative control × 2.1  
 

Positive results: Sample O.D ≥ cut-off O.D  
 

Negative results: Sample O.D < cut-off O.D 
 

2.4 Statistical Analysis 
 

The data obtained from questionnaires 
administered and those obtained from the 
laboratory analysis were managed and analyzed 
using the EPI-INFO version 7.2 statistical 



 
 
 
 

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package. Results were reduced to percentiles 
and a Pearson chi-square test was performed at 
a 95% confidence interval. P-values less than 
0.05 (p< 0.05) was used to determine 
relationships between parameters evaluated and 
seroprevalence.      
 

3. RESULTS AND DISCUSSION 
 

A total of one hundred and eighty-four pregnant 
women attending antenatal clinic at the Federal 
Teaching Hospital Ado Ekiti were enrolled in this 
study. Their ages ranged between 16 to 41 years 
with a mean age of 28±13 years (Table 1).  
 

One hundred and seventy-six (95.6%) of the 
subjects were seropositive for rubella IgG and 22 
(12.0%) for rubella IgM. Twenty (10.9%) were 
seropositive for both rubella IgG and IgM, 2 
(1.1%) only for rubella IgM; and 154 (83.7%) for 
rubella IgG. Seropositivity was higher among age 
group above 35 years: 12 (100%) for IgG and 
age group 31-35 years, 6 (18.8) for IgM (Table 
1). The distribution was however non-statistically 
significant (p = 0.50 and 0.85 respectively). 
 

Higher but non-significant proportion of subjects 
in the third trimester 62 (98.4%) and second 
trimester 84 (96.6%) as against 30 (88.2%) in the 
first trimester were seropositive for IgG. In 
contrast to this pattern, a higher proportion of the 
subjects in first trimester 8 (23.5%) and second 
trimester 12 (13.8%) as against 2 (3.2%) in the 
third trimester were seropositive for rubella IgM. 
The highest rate of co-positivity for rubella IgG 
and IgM was observed in the second trimester 15 
(17.2%) (Table 2). The married ones among the 
subjects had a higher seroprevalence 152 
(97.4%) for rubella IgG while the singles had a 
higher seroprevalence 6 (26.1%) for rubella IgM 
(Table 3). The subjects with no form of formal 
education, zero number of parity and some level 
of knowledge of the virus had the least 
seroprevalence for rubella IgG 3 (60.0%), 53 
(91.3%), 32 (94.1%) respectively. On the other 
hand, those with zero number of parity had 
significant (p = 0.04) seronegativity for rubella 
IgM 56 (93.6%). 
 

Despite the global public health implication of 
Rubella infection which has stimulated the 
endorsement of the Global Measles and Rubella 
Strategic plan in 2012, which was planned to 
cover 2012-2020, it is a thing of concern that 
there is still a paucity of epidemiological data on 
seroprevalence of rubella infection in Nigeria. 
The few that exist are skewed to areas like Zaria, 
Jos, Maiduguri, Abia, and Osogbo. This impact 

negatively on making rightful decisions on 
intervention approaches to curb the incidences of 
CRS.  
 
Rubella IgG serostatus is an indicator of previous 
exposure to the virus or immunization, thus it is 
used as a reliable biomarker for the prevalence 
of the disease where vaccination history is 
known. On the other hand, IgM is an indication of 
a fresh infection and as such used to estimate 
the incidence of the disease. When IgG co-exist 
with IgM, it could be an indicator of an infected 
person who is just recovering and rarely re-
infection. The seronegative population for IgG is 
the susceptible population which is considered 
high risk. 
 
In this present study, 95.6% of the pregnant 
women enrolled were seropositive for anti-rubella 
IgG antibodies. This high seroprevalence is 
similar to 97.9% report by Muhammad et al. [22], 
93.1% by Olajide et al. [5] and 90.2% by Gubio et 
al. [23] all in Zaria, Nigeria. A lower 
seroprevalence was reported from a closer 
location to that of this present study by Kolawole 
et al. [24] who reported 87.5% IgG 
seroprevalence in Osogbo, South-west of 
Nigeria. This lower prevalence is likely due to the 
time lag between the two studies. Lower IgG 
seroprevalences have also been reported by 
Hamdan et al. [25] in Sudan (65.3%) and 
Oyinloye et al. [26] in Maiduguri, Nigeria. This 
indicates variability in the epidemiology of the 
virus from one location to another. 

 
We also recorded 12% seropositivity for anti-
rubella IgM. This was a little higher than the 
findings of Mengouo et al. [27] in Cameroon who 
surveyed 2008 -2014. They, however, record 
12.4% IgM seroprevalence in 2012. The findings 
of Getehun et al. [13] and Junaid et al. [28] were 
higher (39.4 and 45.2% respectively) possibly 
because their studies were not restricted to 
pregnant women and time lag. From this present 
study, 11% of subjects positive for IgM were also 
positive for IgG while 1% was just IgM. This 
could imply that the 12% got infected in the 
recent past but the 11% with IgG and IgM are 
already recovering while the remaining 1% was 
yet to produce IgG (meaning they are still at the 
active stage of infection). Incidentally, the 1% 
population with only IgM were all in the first 
trimester; a high-risk period. The other 12% 
thought to have recovered were all distributed 
through the first, second, and third trimester of 
pregnancy. These imply that most of them 
especially those in the first and early part of the 



 
 
 
 

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Table 1. Seroprevalence of Rubella IgG and IgM based on the age group 
 

 Age group (Years)   
Number 

Rubella IgG Rubella IgM  Rubella IgG and IgM 

Positive (%) p-Value 
 

Positive (%) p-Value 
 

Positive (%) p-Value 
 

16-20 8 5 (62.5)   1 (12.5)   1 (12.5)   
21-25 72 69 (95.8)   8 (11.1)   7 (9.7)   
26-30 60 59 (98.3) 0.50 2.34 6 (10.0) 0.89 0.58 5 (8.3) 0.15 3.76 
31-35 32 31 (96.9)   6 (18.8)   6 (18.8)   
Above 35 12 12 (100)   1 (8.3)   1 (8.3)   
Total 184 176 (95.6)     22 (12.0)     20 (11.0)     

 
Table 2. Seroprevalence of Rubella IgG and IgM in relation to stages of pregnancy 

 

Trimester    
Number 

Rubella IgG Rubella IgM  Rubella IgG and IgM 

Positive (%) p-Value 
 

Positive (%) p-Value 
 

Positive (%) p-Value 
 

First 34 30 (88.2)   8 (23.5)   6(17.6)   
Second 87 84 (96.6) 0.17 2.52 12 (13.8) 0.07 5.12 12 (13.8) 0.06 1.55 
Third 63 62 (98.4)   2 (3.2)    2 (3.2)   
Total 184 176 (95.6)   22 (12.0)   20 (10.9)     

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 



 
 
 
 

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Table 3. Seroprevalence of Rubella IgG and IgM antibodies in relation to some sociodemographic data 
 

Sociodemographic Data   
Number 

Rubella IgG Rubella IgM 

Positive (%) p-Value 
 

Positive (%) p-Value 
 

Marital status       
Married 156 152 (97.4)   15 (9.6)   
Single 23 21 (91.3) 0.76 1.56 6 (26.1) 0.56 0.85 
Separated 5 3 (60.0)   1 (20)   
Educational status        
Primary 52 49 (94.2)   6 (11.5)     
Secondary 86 85 (98.8) 0.10 2.59 10 (11.6) 0.30 0.99 
Tertiary 41 39 (95.1)   5 (12.2)   
None 5 3 (60.0)   1 (20)   
Religion        
Christianity 143 140 (97.9)   17 (11.9)   
Islam 37 34 (91.9) 0.71 0.91 5 (13.5) 0.32 2.12 
Others 4 2 (50.0)   Nil   
Occupation        
Student 15 11 (73.3)   Nil   
Housewife 49 47 (95.9)   8 (16.3)   
Public servant 41 40 (97.6) 0.07 3.10 5 (12.2) 0.07 3.46 
Trader 42 40 (95.2)   6 (14.3)   
Farmer  37 34  (91.9)   3 (8.1)   
Parity       
0 58  53 (91.3)   2 (3.4)   
1-2 79 76 (96.2) 0.05 3.14 14 (18.7) 0.04 3.75 
3-4 29 29 (100)   6 (20.7)   
Above 4 18 18 (100)   Nil   
Knowledge of the virus       
Yes 34 32 (94.1) 0.48 2.01 6 (17.6) 0.42 0.22 
No 150 144 (96.0)   16 (10.7)   



 
 
 
 

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second trimester must have been going through 
the active phase of infection during the first 
trimester which might put the fetus at the risk of 
CRS. The susceptibility of these populations is 
4% going by the IgG seroprevalence, but 
considering that the remaining 11% which had 
both IgG and IgM are likely to have had a recent 
infection and were just recovering; the 
susceptibility could be placed at 15.2%. This is 
comparable with 13.7% susceptibility reported by 
Tamirat et al. [14] in Southern Ethiopia. 
 

Rubella infectivity is not associated with age (p > 
0.05) though ages 16 – 20 had the least 
prevalence for IgG likewise IgM. This agrees with 
the report of Muhammad et al. [22] but contrary 
to some others within the country. This was 
probably due to the high endemic nature of the 
virus in Zaria, Nigeria. The virus is said to be in 
constant circulation among the population thus 
downplaying the effect of some of the risk factors 
like age [22]. A similar situation is observed in 
this present study. Higher rubella IgM 
seroprevalence rates have also been 
documented among children and adolescents 
than in adults with the trend seen to decrease 
with an increase in age [23,28-32]. Antibodies 
were found at every trimester which did not differ 
significantly from one another (p > 0.05), a 
finding that agreed with that of Olajide et al. [5]. 
IgG was highest in the third trimester and IgM 
higher in the First trimester. This is contrary to 
Fokunang et al. [33] who reported the highest 
prevalence in the second trimester. This 
difference may be accrued to cultural variation. 
Some ethnic groups are more predisposed to 
presenting early at antenatal clinics during 
pregnancy than the others. 
 

Formal educational status, religion, and 
occupation had no significant effect (p > 0.05) on 
the distribution of seroprevalence of IgG and IgM 
in this present study. This finding is similar to the 
findings of Mohammed et al. [22]. It was 
observed that though a high percentage of the 
subjects had a tertiary or at least a secondary 
school level formal education, it did not cumulate 
to adequate knowledge about the virus. Even the 
few who indicated knowledge of the virus did not 
request to be vaccinated as all subjects in this 
study claimed not to have been vaccinated 
against the virus. Parity was however associated 
with the risk of rubella infection (p < 0.05). Its 
effect was seen both on IgG and IgM serostatus. 
The nulliparous (zero parity) had a 93.1% IgG 
prevalence and 3.4% for IgM as compared to the 
multiparous that had at least 18.7% and more 
IgM prevalence which aligns with the repost of 

Onakewhor and Chiwuzie [31] but contrary to 
that of Olajide et al. [5] who reported no 
association between reproductive character and 
risk of rubella infection. Parity may be associated 
with the risk of infection since mothers who have 
a number of children may stand a risk of being 
infected from the children [12]. 
 

The study is limited by the unwillingness of some 
of the participants to provide genuine information 
and inability to recall past experiences, hence, 
possibly introducing some biases to the study. 
The accuracy generalizations about pregnant 
women based on this study may also be limited 
because it is an institution based study. A 
subsequent community based longitudinal study 
may resolve these limitations. 
 

4. CONCLUSION 
 

This study reveals a high rate of exposure of 
pregnant women to rubella virus in this study 
area. This suggests continuous transmission and 
endermicity of rubella in the area. Prevalence 
showed similar distribution pattern irrespective of 
sociodemographic features examined. However, 
infection is significant with respect to parity. The 
findings emphasize the need to implement 
routine childhood immunization against rubella 
virus as well as vaccinating susceptible women 
of childbearing age. 
 

CONSENT 
 

The authors declare that written informed 
consent was obtained from the patients for the 
publication of this report. A copy of the written 
consent is available for review by the Editorial 
office/Chief Editor/Editorial Board members of 
this journal. 
 

ETHICAL APPROVAL 
 

The authors hereby declare that Ethical approval 
for the study was obtained from the Ethics and 
Research Committee, Federal Teaching Hospital 
Ido-Ekiti, Reference Number: ERC/2017/02/20/ 
50B. Subjects were fully informed of their right to 
opt-out of the study at any point they so desire. 
They were informed about investigations carried 
out with their samples, potential risk of the study, 
and were assured of confidentiality of the results.  
 

ACKNOWLEDGEMENTS 
 

The authors are grateful to the authority of 
Federal Medical Teaching Hospital Ido-Ekiti who 
permitted us to carry out this study in their facility 
through the ethical committee. The Cooperation 



 
 
 
 

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of the Nurses and Doctors at the antenatal clinic 
during this study is acknowledged. We also 
appreciate Afe Babalola University for granting 
us access to their Medical Laboratory where the 
sample analysis was carried out. 
 

COMPETING INTERESTS 
 
Authors have declared that no competing 
interests exist. 
 

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