Abstract
Typhoid fever is a significant public health burden in low-income countries caused by Salmonella enterica serotype typhi (S.typhi). Clinical manifestations of typhoid fever are varied and non-specific, making the diagnosis difficult. Using oxgall for pre-incubation as a selective culture medium before amplification of Real-time PCR (RT-PCR) in wholeblood produces a fast and sensitive diagnostic. The purpose of this study was to know the performance oxgall-precultured Real-time PCR for detection of Salmonella sp.. Prior to the sample process, spike method optimization was performed to find out that the reagents were well used for clinical specimens. In the sample process, blood samples from 30 Widal-positive patients were collected for this study . Venous blood samples from typhoid fever patients were taken on the day of diagnosis; 5 ml for blood culture, and 5 ml for RT-PCR. The bacteria were grown in oxgall 10% (standard microbiological laboratory clinics) and incubated for 6 hours (37° C) before bacterial DNA was isolated for RT-PCR detection. The results showed that reagen of RT-PCR is good used for a clinical sample and a blood culture was better than RT-PCR using oxgall (positive blood culture results over 24 hours). This suggests that there is a need for further research on the duration of incubation and oxgall concentrations in RT-PCR and the selection of clinical samples.
There is no Figure or data content available for this article
References
2. World Health Organization. Background Document: The Diagnosis, Treatment and Prevention of Typhoid Fever. Geneva: World Health Organization; 2003.
3. Wain J, Hosoglu S. The laboratory diagnosis of enteric fever Review Article The laboratory diagnosis of enteric fever. 2008;(February):10-15. doi:10.3855/jidc.155.
4. Lalith PM, Kumar V, Chanpheaktra N, et al. Typhoid Fever Among Hospitalized Febrile Children in Siem Reap, Cambodia. J J Trop Pediatr. 2012;58(1).
5. Stella IS, Bamidele M, Fowora M, Helen T, Emmanuel. Application of A Point-of-Care Test for The Serodiagnosis of Typhoid Fever in Nigeria and The Need for Improved Diagnoctics. J Infect Dev Ctries. 2011;5(7):520-526.
6. Kundu R, Ganguly N, Ghosh TK, Yewale VN, Shah RC, Shah NK. IAP Task Force Report: Diagnosis of Enteric Fever in Children. Indian Pediatr. 2006;43(10):875–883.
7. Wain J, Pham VB, Ha V, et al. Quantitation of Bacteria in Bone Marrow from Patients with Typhoid Fever: Relationship Between Counts and Clinical Features. J Clin Microbiol. 2001;39(4):1571-1576. doi:0.1128/JCM.39.4.1571–1576.2001.
8. Wain J, Diep TS, Bay PVB, et al. Specimens and Culture Media for The Laboratory Diagnosis of Typhoid Fever. J Infect Dev Ctries. 2008;2(6):469-474. doi:10.3855/jidc.164.
9. Hatta M, Smits HL. Detection of Salmonella Typhi by Nested Polymerase Chain Reaction in Blood, Urine, and Stools Samples. Am J Trop Med Hyg. 2007;76(1):39-143. doi:10.4269/ajtmh.2007.76.139.
10. Ambati SR, Nath G, Das BK. Diagnosis of Typhoid Fever by Polymerase Chain Reaction. Indian J Pediatr. 2007;74(10):909-913.
11. Dorak TM. Real-Time PCR. Newcastle: Taylor and Francis Group; 2006.
12. Julie L, Edwards K, Saunders N. Current Technology and Applications Real-Time PCR. Norfolk: Caister Academic Press; 2009.
13. Ibrahim WA, El-Ghany WA, Nasef SA, Hatem M. Comparative Study On The Use Of Real Time Polymerasechain Reaction (Real-time PCR) And Standard Isolation Techniques For The Detection Of Salmonellae In Broiler Chicks. Int J Vet Sci Med. 2014;2(1):67-71.
14. Siregar TH, Elliman J, Owens L. Development Of Real Time Polymerase Chain Reaction for Detection Salmonella Typhimurium and Salmonella enteriditis in Fish. Squalen. 2012;7(2):51-58.
15. Mufty MM. Application of A Real-time qPCR Method for Detection of Salmonella sp. in Shrimp and Scallop and Its Partial Validation. 2008.
16. Fatimi H. Polymerase Chain Reaction (PCR). Palembang: Program Studi Ilmu Biomedik, Program Pasca Sarjana, Universitas Brawijaya; 2010.
17. Zhou L, Jones C, Gibani MM, et al. Development and Evaluation of a Blood Culture PCR Assay for Rapid Detection of Salmonella Paratyphi A in Clinical Samples. PLoS One. 2016. doi:10.1371/journal.pone.0150576.
18. Colemen W, Buxton BH. The Bacteriology of The Blood in Convalescence from Typhoid Fever with A Theory of The Pathogenesis of The Disease. J Med Res. 1909;21(1):83-93.
19. Kaye D, Palmieri M, Rocha H. Effect of Bile on the Action of Blood Against Salmonella. J Bacteriol. 1966;91(3):945-952.
20. Zhou L, Pollard AJ. A Novel Method of Selective Removal of Human DNA Improves PCR Sensitivity for Detection of Salmonella Typhi in Blood Samples. BioMed Cent Infect Dis. 2012. doi:10.1186/1471-2334-12-164.
21. Prakash P, Mishra P, Singh AK, Gulati AK, Nath G. Evaluation of Nested PCR in Diagnosis of Typhoid Fever. J Clin Microbiol. 2005;43(1):431-432. doi:0.1128/JCM.43.1.431-432.2005.
22. Konemann EW, Allen SD, Janda WM, Schreckenberger PC, Winn WC. Color Atlas and Textbook of Diagnostic Microbiology. 5th ed. Philladelpia: Lippincott Williams and Wilkins; 1997.
23. Bhan MK, Bahl R, Bhatnagar S. Typhoid and Paratyphoid Fever. Lancet. 2005:749-762. doi:10.1016/S0140-6736(05)67181-4.
24. Andualem G, Abebe T, Kebede N, Gebre-Selassie S, Mihret A, Alemayehu H. A Comparative Study of Widal Test with Blood Culture in The Diagnosis of Typhoid Fever in Febrile Patients. BioMed Cent Res Notes. 2014. doi:10.1186/1756-0500-7-653.
25. Ernawati. Perbandingan positivitas pemeriksaan Real-time PCR(REAL TIME PCR) Gen invA Salmonella typhi Dari Bahan Biologis Tersimpan Buffycoat dengan Pemeriksaan Kultur dari Sampel Darah Segar untuk Mediagnosis Demam Tifoid. 2016.
How to Cite This
Article Metrics
Download Statistics
Downloads
Other Statistics
Copyright and Permissions
Publishing your paper with Jurnal Teknologi Laboratorium (JTL) means that the author or authors retain the copyright in the paper. JTL granted an exclusive reuse license by the author(s), but the author(s) are able to put the paper onto a website, distribute it to colleagues, give it to students, use it in your thesis etc, even commercially. The author(s) can reuse the figures and tables and other information contained in their paper published by JTL in future papers or work without having to ask anyone for permission, provided that the figures, tables or other information that is included in the new paper or work properly references the published paper as the source of the figures, tables or other information, and the new paper or work is not direct at private monetary gain or commercial advantage.
JTL journal provides immediate open access to its content on the principle that making research freely available to the public supports a greater global exchange of knowledge. This journal is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License. This license lets others remix, transform, and build upon the material for any purpose, even commercially.
JTL journal Open Access articles are distributed under this Creative Commons Attribution-ShareAlike 4.0 International License (CC BY-SA). Articles can be read and shared for All purposes under the following conditions:
- BY: You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.
- SA: If you remix, transform, or build upon the material, you must distribute your contributions under the same license as the original.