Rotavirus infection among children under 5 years ...

‫مجلة النيلين الطبية – العدد التاسع‬

NMJ March 2013

Rotavirus infection among children under 5 years presenting with diarrhea to Ahmed Gasim pediatric hospital, Khartoum

Kafi SK 1* , Osman MM 2, Musa HA 3 1: Shamsoun Khamis Kafi: Microbiology Department, faculty of medical laboratory science, The National Ribat University, Khartoum, Sudan. 2: Mahasin Mohamed Osman Said Ahamed, Department of Microbiology, Alzaeem Alazhari University, Khartoum North, Sudan. 3: Hassan Abdelaziz Musa, Department of Microbiology, Faculty of Medicine, The National Ribat University, Khartoum, Sudan.

* corresponding author : [email protected]

ABSTRACT Background: Rotavirus is the most common cause of viral diarrhea worldwide killing up to 850000 children annually. WHO reported that 20 - 70% of all hospitalization and 20% of deaths from diarrhea were attributable to rotavirus. Few studies were conducted in Sudan to study rotavirus infection. The current study is conducted to determine the role of rotavirus in the etiology of diarrhea among children presenting to Ahmed Gasim pediatric hospital. Methodology: During the period from June,2009 to May, 2010, stool specimens were collected from 446 children under 5 years presenting to Ahmed Gasim hospital with diarrhea. At the same time, 5mls of venous blood were collected from each patient under study. The stool specimens were tested for Rotavirus antigen by enzyme immunoassay kit. The blood specimens were analyzed for urea and electrolytes in addition to full blood count. Results: One third (33%) of the children were found to be positive for rotavirus infection. The highest rate of infection (33.7%) was in the age group 6 - 12 months. Rotavirus infection was found to occur all over the year but higher in November to January. Hyponatraemia and hypokalaemia were the most frequent biochemical changes in children infected with rotavirus. Lymphocytosis(83%) and anemia(71%) were the commonest 6  2013 Al Neelain Medical Journal

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hematological abnormalities. The rate of infection with rotavirus obtained in this study (33%)is high but comparable with the reported figures from other countries. Rotavirus infection was found exist throughout the year but higher in the Winter months; this is in agreement with previous studies. The most frequent biochemical changes found in this study(hyponataemia and hypokalaemia) are expected in a patient suffering from diarrhea. Lymphocytosis is also relevant to virus infection. Conclusion: Rotavirus virus was found to play considerable role in the etiology of diarrhea in children below the age of 5 years in Ahmed Gasim hospital.

INTRODUCTION

Rotavirus is the most common cause of

infection to severe and on occasion fatal

severe dehydrating gastroenteritis among

dehydrating illness. Typically the clinical

children globally, resulting in approximately

presentation is of 3 days vomiting and 5

600,000 to 850,000 deaths each year. Most

days of watery diarrhea with moderate fever

deaths occur in developing countries, where

following a 2 days incubation period.(4,5)

access to rehydration therapy and other

Despite these facts, the role of

medical care is often limited. The disease

rotavirus in the etiology of diarrheal diseases

burden is also unlikely to be significantly

had not thoroughly been studied in Sudan

reduced by improvements in hygiene and

evidenced by lack of reported studies on the

sanitation. A

(1,2)

.

WHO-sponsored

subject. The aim of this study was to review

of

estimate the rate of infection with Rotavirus

rotavirus studies found that 20–70% of all

among children below 5 years presenting to

hospitalizations and 20% of deaths from

the study area with Gastroenteritis and to

diarrhea were attributable to rotavirus(3).

determine hematological and chemical

Rota virus infection may produce a spectrum

changes caused by the virus.

of illnesses ranging from sub clinical

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METHODOLOGY The study was conducted during the period

All the collected stools specimens were

from

tested for rotavirus antigen by means of a

June 2009 to May 2010 in Ahmed

Gasim pediatric hospital to determine the

commercial

rate of infection with rotavirus among

(IDEIATM Rotavirus

children below 5 years presenting with

IDEIA

gastroenteritis

polyclonal

and

to

determine

the

TM

enzyme

immunoassay REF/K60200.

Rotavirus antibody

test in

a

utilizes

kit The a

solid-phase

hematological and biochemical changes

sandwich enzyme immunoassay to detect

which occur as a result of infection.

group

antigen

present

in

Group

A

rotaviruses. Break- apart micro wells are

The purpose of the study was first explained

coated with a rotavirus specific polyclonal

to both or any of the parents' and after

antibody.

agreeing to enroll the child in the study; asked to fill a questionnaire. Requested

The test procedure was as follows: All

information included the demographic data

reagents and stool specimens were allowed

and the clinical symptoms and signs.

to reach the room temperature (15- 30oC)

Thereafter, approximately, 5 mls of stool

before use.100l of the negative control,

were collected from each child under study

positive control and each specimen were

in sterile, wide mouth, leak proof plastic

added to the corresponding micro wells of

container. The collected specimens were

the micro titer plate(2 wells for the negative

₀

stored in a deep freezer at -20 C until tested.

and one for the positive controls).The plate

Care was taken to avoid repeated thawing

was incubated at 20-30 oC for 60 minutes.

and freezing cycles.

Then washed 5 times using wash solution. 100l of the Conjugate were added to each

In addition 5mls of venous blood were

micro well and the plate incubated for 10

collected from each subject under study

minutes at 20 - 30 C. The plate was washed

under aseptic technique. Of the total volume

again 5 times as above. 100L of the

of blood, 2mls were added to EDTA

substrate were added resulting in immediate

containers and the remaining 3 mls into

development of blue color. Thereafter

heparin container. 8  2013 Al Neelain Medical Journal

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100µL of stop solution were added changing

Biochemical analysis:

the color into yellow.

The heparinized blood samples were used

-The absorbance was read photo metrically

for estimation of blood urea and electrolytes

at 450nm.

as follows:

Validation of the test:

a)Estimation of Urea ( Berthelot enzymatic method):

The test was considered valid and accepted, if the

absorbance of the positive control

The blood urea level was estimated

micro well was greater than 0.500 and the

using Berthelot enzymatic method

mean absorbance of the negative controls

b) Estimation of electrolytes:

less than 0.150. Otherwise the test was

Potassium and Sodium electrolytes were

considered invalid and repeated.

measured by emission spectrometry using a Calculation of the cut-off value:

flame photometer.

The cut-off value was calculated by

Hematological analysis:

adding 0.100 to the mean absorbance of the

This was done using EDTA blood samples.

negative controls.

Complete blood picture was done using Interpretation of the results: Specimens with the absorbance ≥

hematological cutoff

hemoglobin,

analyzer total

white

to

estimate

blood

cells,

value were considered positive for rotavirus

differential (neutrophil, lymphocyte and

antigen.

other white blood cells).

RESULTS A total of 446 children were included in this

of the studied children were males and

study. The majority were below the age of

almost 50% were from Gabel Awalia

two years(table:1). Almost two thirds (64%)

locality. 9

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Among the studied children, 146

throughout the year but higher in winter

(33%) were found to be positive for

(November to January) (Figure:1).

rotavirus infection. The highest rate of

Biochemical findings:

infection was in the age group 6-12 months

Hyponatraemia

(table: 1).

frequent biochemical change in Rotavirus

revealed the

infected children(Table:3). Results of blood

highest rate of infection 36.4%(135/371)

urea showed that 22% have low urea level

among patients residing in the peripheral

and 29% high urea level (table:3).

localities(Sharg elneel, Ombada, Karary and

Hematological changes:

Gabal Awalia) collectively compared to those residing in the central Localities( Khartoum,

Khartoum

North

About 71% of the infected children

and

were

Omdurman) with infection rate of 14.7% (

found

to

have

anemia,

83%

lymphocytosis and 54% leucocytosis (tables

table :2).

4).

Seasonal variation: Attempt

hypokalemia

(67% & 73% respectively) were the most

Distribution of rotavirus positive cases according to locality

and

to

Among show

the

the

positive

cases

of

Rotavirus 7% were found to be treated with

seasonal

ant malarial drugs.

variation in the rate of infection by Rotavirus revealed that infection occurs


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Table 1 : Age distribution of the rotavirus infected children(146). Age group

Total tested

Rotavirus + ve Freq.

Percent

Less than 6 months.

94

30

31.0

6 - 12 months.

231

78

33.7

13 - 18 months.

73

24

32.9

19 - 24 months

38

12

31.6

25months to 5years.

10

2

20

Total

446

146

33

Table 2: Distribution of Rotavirus positive Cases according to residence Locality

Total tested

Rotavirus +ve Freq.

Percent

Khartoum North#

5

2

40

Sharg Elneel*

70

24

34

Omdurman#

23

4

17

Ombada*

13

5

38

Karary*

3

1

33

Khartoum #

47

5

11

Gabal Awalia*

223

87

39

Other States

62

18

29

Total.

446

146

33

key:* = peripheral locality

# = central locality 11

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Table(3): Biochemical changes in the Rotavirus infected children(146) Parameter

Urea Percent 28.8

Freq. 0

Sodium Percent 0

Freq. 3

72

49.3

48

32.9

36

24.7

32

21.9

98

67.1

107

73.3

Above the normal range Within the normal range Below the normal range Reference values

Freq. 42

18- 45 mg/dl

135-150 mmol/L

Potassium Percent 2.1

3.5 – 5 mmol/L

Table(4): Hematological changes in the Rotavirus infected group(146). Parameter

Within normal range Below normal range Above normal range Normal range

TWBC/Cmm

Neutrophils Cmm Freq. % 109 74.7

Lymphocyte Cmm Freq. % 25 17.1

Freq. 49

% 33.6

0.0

0.0

12

8.2

0.0

97

66.4

25

17.1

121

4000 - 10000

2000 - 7000

Hemoglobin(gm/dl) Freq. 42

% 28.8

0.0

104

71.2

82.9

0.0

0.0

1000 - 3000

12 - 16


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DISCUSSION In the current study, rotavirus was found to

conducted studies in Vietnam, Netherland

be responsible for diarrhea in children

and Newzeland (9,10, 11).

admitted to Ahmed Gasim hospital during

The highest rate of infection with

the study period. This finding is in concordance

with

previous

rotavirus was found to be within the age

studies

group 6 - 12 months followed by 13 - 24

conducted in Malawie and Taiwan that

months and males were more affected than

reported prevalence of 34% and 34.6% respectively(6,

7)

females. These findings are in agreement

. WHO also reported a

with

prevalence of 34% in 1985(8). The rate of

previous

studies

conducted

in

Sweden(9).

rotavirus infection obtained in this study was

Regarding the seasonal variation of

however lower than in other previously

rotavirus infection, this study like other 13  2013 Al Neelain Medical Journal

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previous studies reported September and

Neutrophils count was within the

December(winter) to reveal the highest rate

normal limits in the majority of rotavirus

of infection with rotavirus(12).

infected children but lymphocytosis was the most prominent hematological abnormality.

Hyponatremia and hypokalemia(67%

These findings are consistent with virus

, 78% respectively) were the most frequent

infection.

biochemical finding in the studied group and both be explained by sodium and potassium loss due to diarrhea.

CONCLUSION Rotavirus is responsible for 33% of hospital admission in

children under 5

Based on these findings, vaccination

years

against

rotavirus

is

recommended

in

presenting with diarrhea. Males are more

addition to further in depth studies to

affected than females. Rotavirus infection is

determine the serotypes responsible for

commoner during Winter (September to

diarrhea in Sudan.

December). ACKNOWLEDGEMENTS: The authors would like to thank Dr. Mobarak Karsany and Mrs. Rehab Elagib for their great help in facilitating the laboratory work. REFERENCES 1- Kosek M, Bern C, Guerrant R

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