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ORIGINAL ARTICLE
Year : 2014  |  Volume : 41  |  Issue : 2  |  Page : 114-117

A comparative clinical study of efficacy of microimmuno assay with WIDAL-test in enteric fever in children


1 Department of Pediatrics, and Microbiology, KIMS Hospital, Bengaluru, India
2 Department of Microbiology, KIMS Hospital, Bengaluru, Karnataka, India
3 Biostatistician, National Institute of Veterinary Epidemiology and Disease Informatics (NIVEDI), Bengaluru, Karnataka, India

Date of Web Publication20-May-2014

Correspondence Address:
Gandarajapura Nagaraj Madhu
#15, "Manjushri", 1st Cross, 1st Main, Sundarnagar, Gokula, Bengaluru - 560 054, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0974-5009.132856

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  Abstract 

The diagnosis of typhoid fever in young children is also a dilemma because of its manifestations and typical presentation may not be seen in all cases. Antibodies to Salmonella typhi antigen are developed in the human body, which can be detected as a diagnostic test for the enteric fever. Objective: This study was undertaken to compare the efficacy of WIDAL-test with micro-immunoassay (dot enzyme immunosorbent assay). Method: 40 cases of clinically suspected enteric fever cases were included in this study. Result: In the present study, nearly 92% were positive for micro immunoassay (dot-enzyme immunosorbent assay) by Enterocheck-WB kit, 80% were positive for WIDAL and only 15% were culture positive. Immunoassay positive, but WIDAL negative cases were 20%, whereas WIDAL positive and immunoassay negative cases were only 7.5%. The positive predictivity of micro-immunoassay in diagnosing enteric fever is better than WIDAL both in 1st and 2nd week of illness. Micro-immunoassay done in the study was rapid in diagnosing the case. Conclusion: It is concluded from the present study that the micro-immunoassay (Enterocheck-WB) is better than WIDAL-test in the diagnosis of enteric fever in children.

Keywords: Enteric fever, Enterocheck-WB kit, micro-immunoassay, WIDAL-test


How to cite this article:
Madhu GN, Srinivasa S, Ravikumar KL, Suresh KP. A comparative clinical study of efficacy of microimmuno assay with WIDAL-test in enteric fever in children. J Sci Soc 2014;41:114-7

How to cite this URL:
Madhu GN, Srinivasa S, Ravikumar KL, Suresh KP. A comparative clinical study of efficacy of microimmuno assay with WIDAL-test in enteric fever in children. J Sci Soc [serial online] 2014 [cited 2020 Aug 12];41:114-7. Available from: http://www.jscisociety.com/text.asp?2014/41/2/114/132856


  Introduction Top


Enteric fever or typhoid fever is a severe systemic disease that is found mainly in developing countries, but it is encountered world-wide because of international travel. The diagnosis of typhoid fever in young children is also a dilemma because of its manifestations and typical presentation may not be seen in all cases. Isolation of Salmonella typhi organisms either by blood culture or cultures of bone marrow, stool and urine or bile juice is considered as the gold standard. Due to parent's urgency and enthusiastic doctors, children receive antibiotics prior to diagnosis which makes the isolation of the S. typhi organisms difficult. Antibodies to S. typhi antigen are developed in the human body, which can be detected as a diagnostic test for the enteric fever.

In WIDAL-test the agglutination titer will depend on the stage of disease. Agglutinins will usually appear by the end of the 1 st week, so that blood taken earlier may give a negative result. The titer increases steadily until the third or the 4 th week, after which it declines gradually. [1] Complexity and higher costs of molecular tests hinder its application in routine use. [2]

A case or carrier is infectious as long as bacilli appears in the stool or urine. [3] Because of intermittent and low level bacteremia, repeated blood cultures should be obtained in suspected cases. [4] Complications can be reduced with timely accurate diagnosis and with adequate chemotherapy [5],[6] Clinical suspicion is necessary before the laboratory assistance can be mobilized. [7]

This study was undertaken to compare the efficacy of WIDAL-test with micro-immunoassay (dot enzyme immunosorbent assay).

Objective

The objective of this study is to compare the micro immunoassay-IgM detection (dot enzyme immunosorbent assay) with WIDAL-test.


  Materials and methods Top


Source of data

The present study was conducted at Kempegowda Institute of Medical Sciences Hospital over a period of 1΍ year (June 2009 to December 2010). In this prospective clinical study, suspected cases of enteric fever up to fifteen were admitted to the pediatric ward.

Inclusion criteria

Clinically suspected cases of enteric fever up to the age of 15 years.

Exclusion criteria

1. Patients with enteric fever above the age of 15 years.

2. Patients who have other associated conditions.

On admission, these subjects were enrolled in to the study. A detailed history was taken, clinical examination was done. A detailed questionnaire was recorded as per the proforma. These subjects were investigated for Hb%, total count, differential count, erythrocyte sedimentation rate, blood culture, WIDAL-test, micro immunoassay-IgM detection, stool routine, urine routine.

Blood sample for culture was drawn and inoculated in to blood culture bottles (before starting antibiotics) and sent to the laboratory, where the culture was done using biphasic brain heart infusion agar.

WIDAL-test by slide and tube agglutination technique, which was expressed in dilution titers of 1/20, 1/40, 1/80, 1/160, 1/320 and 1/640. Another serological test, micro immunoassay (dot enzyme immunosorbent assay) was done with Enterocheck-WB kit to detect IgM class of antibodies directed to lipopolysaccharide (LPS) 'O' of S. typhi organism and expressed as present or absent.

The dot-enzyme immunoassay (EIA) is a relatively newer serologic test based upon the presence of specific IgG and IgM antibodies to a specific 50-kD outer membrane protein (OMP) antigen on the S. typhi strain. The test incorporates nitrocellulose strips impregnated with the OMP antigen and separately identifies IgG and IgM antibodies. [1]

The dot enzyme immunosorbent assay allows separate visual assessment of the presence of specific IgG and IgM antibodies to the OMP in a standard 1:100 dilution of serum by a characteristic color change and has been reported to be at least as specific and sensitive as WIDAL-test in children with typhoid fever. [8] The cases, which were negative for blood cultures, WIDAL and micro immunoassay (dot-enzyme immunosorbent assay by Enterocheck-WB) even in the 2 nd week of illness, were excluded from the study.

Sample size: 40 cases.


  Results Top


In the present study, fever was the symptom present in all the cases (100%).

In the present study, 11 cases out of 40 cases (27.5%) presented in the 1 st week, 28 cases presented in the 2 nd week (70%), only one case presented in the 3 rd week of fever (2.5%).

Vomiting was noticed in 11 cases (27.5%).

Pain abdomen was the symptom in 15% of the cases.

Loose stools were present in 2 cases (5%).

Cough was present in 7 cases i.e., 17.5%.

In the present study 37 cases out of 40 (92%) were positive for micro immunoassay by Enterocheck-WB kit, 32 cases (80%) were positive for WIDAL and 6 cases (15%) were culture positive. Out of 11 cases admitted in the 1 st week of fever immunoassay + and WIDAL - were 6 cases, immunoassay + and WIDAL + were 3 cases, immunoassay - and WIDAL + were 2 cases, blood culture + and immunoassay + were 5 cases, blood culture + WIDAL + were 2 cases, blood culture +, WIDAL + and immunoassay + were 2 cases. Out of 28 cases admitted in the 2 nd week of fever immunoassay + and WIDAL - were 2 cases, immunoassay + and WIDAL + were 25 cases, immunoassay - and WIDAL + were 1 case, blood culture + and immunoassay + were 1 case, blood culture + WIDAL + were 1 case, blood culture +, WIDAL + and immunoassay + were 1 case as shown in Graph 1 [Additional file 1] and Graph 2 [Additional file 2].

Only one case was admitted in the 3 rd week of fever, where both immunoassay and WIDAL was positive.

[Table 1] shows, in the present study, out of 11 cases presented in the 1 st week of fever: 6 cases were positive by immunoassay and were negative for WIDAL, 2 cases were WIDAL positive an immunoassay negative and in 3 cases both immunoassay and WIDAL were positive. Out of 28 cases presented in the 2 nd week of fever: 2 cases were positive by immuno assay and were negative for WIDAL, 1 case was WIDAL positive and immunoassay negative and in 25 cases both immunoassay and WIDAL were positive. Only one case presented in the 3 rd week of fever that was positive by both WIDAL and micro immunoassay.
Table 1: Comparison of investigation methods (WIDAL and micro immunoassay)[9]

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The sensitivity of WIDAL-test is 78.4% and the sensitivity of micro immunoassay is 90.6% in the present study [Table 2] and Graph 3 [Additional file 3].
Table 2: Vise-versa comparison of positive with negative tests of WIDAL and micro immunoassay[9]


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  Discussion Top


In the present study, 37 cases out of 40 (92%) were positive for micro immunoassay (dot-enzyme immunosorbent assay) by Enterocheck-WB kit, 32 cases (80%) were positive for WIDAL and 6 cases (15%) were culture positive. Zulfiqar Ahmed Bhutta et al. reported 52 cases out of 71 (73%) in the entire cohort and 39 cases out of 46 cases (84%) among culture proven typhoid fever cases were positive by EIA (Typhidot-M). 39 cases (54%) in the entire cohort and 29 (63%) cases among culture proven typhoid fever cases were positive for WIDAL-test. [8]

Deborah House et al. reported serological assays based on the detection of IgM antibodies against either serotype Typhi LPS (ELISA) or whole bacteria (dipstick) had a significantly higher sensitivity than the WIDAL TO test when used with a single acute phase serum sample (pd"0.007). [10] Mary V Jesudasan et al. reported that detection of anti LPS antibodies by ELISA had a sensitivity of 89.4% and specificity of 94.9% in their study. [11]


  Conclusion Top


In the present study, 92% were positive for micro immunoassay (dot-enzyme immunosorbent assay) by Enterocheck-WB kit, 80% were positive for WIDAL and only 15% were culture positive. Immunoassay positive but WIDAL negative cases were 20% whereas WIDAL positive and immunoassay negative cases were only 7.5%. The positive predictivity of micro-mmunoassay in diagnosing enteric fever is better than WIDAL both in 1 st and 2 nd week of illness.

Its concluded from the present study that the micro immunoassay (by Enterocheck-WB) is better than WIDAL-test in the diagnosis of enteric fever in children. Micro-immunoassay done in the study was rapid in diagnosing the case, comparatively economical and can be done on the out-patient basis with a small quantity of serum or whole blood (5 μl).

 
  References Top

1.Balakrishna TP, Sumathi S, Anuradha K, Venkatesh D, Krishna S, et al. A comparative study of typhidot and Widal test in the diagnosis of typhoid fever. J Evol Med Dent Sci 2013;2:3721.  Back to cited text no. 1
    
2.Pastoor R, Hatta M, Abdoel TH, Smits HL. Simple, rapid, and affordable point-of-care test for the serodiagnosis of typhoid fever. Diagn Microbiol Infect Dis 2008;61:129-34.  Back to cited text no. 2
    
3.Park K. Park′s Text Book of Preventive and Social Medicine. 18 th ed. Jabalpur: M/S Banarsidas Bhanot; 2005. p. 195-8.  Back to cited text no. 3
    
4.Behrman RE, Kliegman RM, Jenson HB. Nelson Text Book of Pediatrics. 17 th ed. Philadelphia: WB Saunders Company; 2004. p. 916-8.  Back to cited text no. 4
    
5.Collier L, Balow A, Sussman M, Duerden BI. Topley and Willson′s Microbiology and Microbial Infections. Systemic Bacteriology. 9 th ed. Vol. 2. London: Arnod; 1998. p. 969-95.  Back to cited text no. 5
    
6.Rator MY. Typhoid fever. Indian J Pract Doct 2004;1:1.  Back to cited text no. 6
    
7.Olsen SJ, Pruckler J, Bibb W, Nguyen TM, Tran MT, Nguyen TM, et al. Evaluation of rapid diagnostic tests for typhoid fever. J Clin Microbiol 2004;42:1885-9.  Back to cited text no. 7
    
8.Sherwal BL, Dhamija RK, Radhawa VS, Jais M, Kaintura A, Kumar M. A comparative study of typhidot and Widal test in patients of typhoid fever. J Indian Acad Clin Med 2004;5:224-6.  Back to cited text no. 8
    
9.Madhu GN. "A comparative clinical study of efficacy of short course ofloxacin therapy with ceftriaxone in uncomplicated enteric fever in children and efficacy of micro immuno assay with widal test." Unpublished data of Rajiv Gandhi University of Health Sciences (RGUHS).  Back to cited text no. 9
    
10.Agrawal PK, Gogia A, Gupta RK. Typhoid fever. J Indian Acad Clin Med 2004;5:60-4.  Back to cited text no. 10
    
11.Dutta S, Sur D, Manna B, Sen B, Deb AK, Deen JL, et al. Evaluation of new-generation serologic tests for the diagnosis of typhoid fever: Data from a community-based surveillance in Calcutta, India. Diagn Microbiol Infect Dis 2006;56:359-65.  Back to cited text no. 11
    



 
 
    Tables

  [Table 1], [Table 2]



 

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