Evaluation of a Rapid Serological Test for Diagnosis of Typhoid Fever Detecting Anti-O9 Antibodies Thesis Submitted for Partial Fulfillment of Master Degree in Medical Microbiology & Immunology Presented by Ahmad Mohammad Al-Shanawany M.B.B.CH Faculty of Medicine Tanta University Under Supervision of Prof. Dr. Tahany Ahmad Abdel- Raouf Professor of Medical Microbiology & Immunology Faculty of Medicine Ain-Shams University Dr. Rasha Ahmad Reda Nasr Assistant Professor of Medical Microbiology & Immunology Faculty of Medicine Ain-Shams University Faculty of Medicine Ain Shams University Faculty of Medicine Medical Microbiology & Immunology Dep.
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Evaluation of a Rapid Serological Test for Diagnosis of Typhoid Fever Detecting Anti-O9 Antibodies
Thesis
Submitted for Partial Fulfillment of Master Degree in
Medical Microbiology & Immunology
Presented by Ahmad Mohammad Al-Shanawany
M.B.B.CH
Faculty of Medicine Tanta University
Under Supervision of
Prof. Dr. Tahany Ahmad Abdel-
Raouf
Professor of Medical Microbiology & Immunology
Faculty of Medicine
Ain-Shams University
Dr. Rasha Ahmad Reda Nasr
Assistant Professor of Medical Microbiology & Immunology
Faculty of Medicine
Ain-Shams University
Faculty of Medicine
Ain Shams University
Faculty of Medicine
Medical Microbiology & Immunology Dep.
Evaluation of a Rapid Serological Test for Evaluation of a Rapid Serological Test for Evaluation of a Rapid Serological Test for Evaluation of a Rapid Serological Test for
Diagnosis of Typhoid Fever Detecting Diagnosis of Typhoid Fever Detecting Diagnosis of Typhoid Fever Detecting Diagnosis of Typhoid Fever Detecting
Protocol of ThesisProtocol of ThesisProtocol of ThesisProtocol of Thesis
Submitted for Partial Fulfillment of Master
Degree in Medical Microbiology& Immunology
Presented byPresented byPresented byPresented by Ahmad Mohammad Al-Shanawany
M.B.B.M.B.B.M.B.B.M.B.B.CHCHCHCH
Faculty of Medicine Faculty of Medicine Faculty of Medicine Faculty of Medicine TantaTantaTantaTanta University University University University
Supervised bySupervised bySupervised bySupervised by
Prof. Dr. Tahany Prof. Dr. Tahany Prof. Dr. Tahany Prof. Dr. Tahany AhmAhmAhmAhmaaaad d d d AbdelAbdelAbdelAbdel----RaoufRaoufRaoufRaouf Professor of Professor of Professor of Professor of Medical Medical Medical Medical Microbiology & ImmunologyMicrobiology & ImmunologyMicrobiology & ImmunologyMicrobiology & Immunology
Faculty of Medicine Faculty of Medicine Faculty of Medicine Faculty of Medicine
AinAinAinAin----Shams UniversityShams UniversityShams UniversityShams University
Dr. Rasha AhmDr. Rasha AhmDr. Rasha AhmDr. Rasha Ahmaaaad Reda Nasrd Reda Nasrd Reda Nasrd Reda Nasr
Assistant professor of Assistant professor of Assistant professor of Assistant professor of Medical Medical Medical Medical Microbiology & ImMicrobiology & ImMicrobiology & ImMicrobiology & Immunologymunologymunologymunology
Faculty of Medicine Faculty of Medicine Faculty of Medicine Faculty of Medicine
AinAinAinAin----Shams UniversityShams UniversityShams UniversityShams University
Faculty of Medicine
Ain-Shams University
2008
Introduction
Typhoid fever is a febrile and often serious systemic illness caused
by Salmonella enterica typhi (S.typhi). It remains a global threat with an
estimated 21 million illnesses and more than 200 000 deaths annually
(Crump et al., 2004). Rapid accurate diagnosis and early treatment with
suitable antimicrobials is essential for rapid recovery and for prevention of
complications and deaths due to this disease and also for the control of
transmission (WHO, 2003).
Because the clinical features of typhoid fever are nonspecific, the
isolation of the causative organism from blood, bone marrow, or stool is
required to confirm diagnosis (Duta et al., 2006). However, the problem
with culture is that the method is laborious and time consuming (taking
days) and even stool cultures are positive in only 30% of patient with acute
illness (Parry et al., 2002).
Serology based on antibody detection has been widely used to
diagnose typhoid fever since the introduction of the Widal test (Widal,
1896). This test has remained the mainstay of serological test in many of
the endemic countries despite numerous accounts of its shortcomings
(Quiroga et al., 1992, Parry et al., 1999 and Nuseba et al., 2002). The
main reason for its continued use is the fact that it is simple-requiring just a
single step with no instrumentation and is affordable by the affected
countries (Tam et al., 2008).
Several new serological tests for typhoid fever have been introduced
which detect antibodies to various purified antigens of S. typhi (Rahman et
al., 2007). Of these tests is the TUBEX O9 antibody test. In this test, when
magnetic particles coated with S. typhi LPS are mixed with indicator
(colored) particles coated with a monoclonal antibody specific for the O9
antigen in S. typhi lipopolysaccharide (LPS), the two types of particles will
bind together and sediment by use of magnet leaving a clear supernatant. If
a patient’s anti-O9 antibodies are present in the reaction mixture, they will
inhibit the binding of the colored particles to the magnetic particles.
Consequently, the supernatant remains colored, the results are graded from
0 (pink, most negative) to 10 (blue, more positive) according to the color
chart provided (Oracz et al., 2003).
Aim of the Work
The aim of this study is to evaluate a rapid serological test in
diagnosis of typhoid fever by detecting anti-O9 antibodies of S.typhi in
comparison with isolation of S.typhi and Widal test.
Patients and Methods
Patients
This study will include 30 patients selected from Armed Forces
Tropical Hospital in Almaza, Cairo, presenting by clinical suspicion of
typhoid fever. All patients will be subjected to full history taking and
clinical examination.
Methods
A- Blood sample will be taken from each patient to do the following:
� Blood culture for isolation & identification of S. typhi.
� Widal test for detection of salmonella antibodies in the patients' serum.
� Detection of IgM antibodies specific for S. typhi O9 LPS by TUBEX O9
antibody test (IDL Biotech, Sweden).
B- Stool samples will be collected for isolation and identification of
S. typhi.
References
• Crump JA, Luby SP and Mintz ED. (2004): The global burden of
typhoid fever. Bull WHO 82: 346-352
• Dutta J, Sur D, Manna B, Sen B, Deb AK, Deen JL, Wain J, Seidlein
LV, Ochiai L, Clemens JD and Bhattachary SK. (2006):
Evaluation of new-generation serologic tests for the diagnosis of