In-house Loop Amplification (LAMP) method for the diagnosis of Salmonella Paratyphi A in low-resource settings
| Autor: | H. Azura, I. Asma, M. Maizan, Ismail Aziah, A. Julia, D. Zawiyah, S. Faizul-Rahman, AR Zaidah, S. Nur Eliana |
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| Rok vydání: | 2014 |
| Předmět: |
Microbiology (medical)
Serotype Salmonella medicine.diagnostic_test Serial dilution Paratyphoid fever Salmonella paratyphi A Gold standard (test) Biology medicine.disease medicine.disease_cause biology.organism_classification complex mixtures Virology Microbiology fluids and secretions Infectious Diseases medicine bacteria Blood culture Bacteria |
| Zdroj: | Asian Pacific Journal of Tropical Disease. 4:251 |
| ISSN: | 2222-1808 |
| DOI: | 10.1016/s2222-1808(14)60567-3 |
| Popis: | Introduction Salmonella Paratyphi A (S. Paratyphi A) causes paratyphoid fever, a disease endemic in developing countries. It is transmitted via the faecal-oral route, mainly through contaminated food and water. The prevalence or incidence of the disease is increasing especially in China, India and Asia. Current diagnosis of S. Paratyphi A is via culture and molecular methods such as PCR. However, these methods do not offer a rapid, simple, and cost-effective detection of S. Paratyphi A especially for use in resource-limited settings. Objective The study was aimed at developing an in-house LAMP method as a rapid, sensitive, specific and cost-effective detection of S. Paratyphi A. Methods An In-house LAMP method was developed and optimized for the detection of S. Paratyphi A using primers that were designed based on intergenic region of SSPA1723a and SSPA1723 gene of S. Paratyphi A. The primers' specificity was tested on 60 bacteria strains consisting of 25 S. Paratyphi A, 20 other Salmonella serovars, and 15 non-Salmonella bacteria which were obtained from Institute for Research in Molecular Medicine (INFORMM) culture bank. Detection limit of LAMP was determined using 10-fold serial dilutions of S. Paratyphi A strain DNA and compared with culture and PCR results. The assay was further evaluated on 60 BACTEC blood culture broths suspected of S. Paratyphi A. The results were compared with those obtained by PCR assay and culture method. Results & Discussion In-house LAMP method was successfully established and optimized. The sensitivity of the assay was determined at 200 CFU. This LAMP assay gave positive result to all the 25 S. Paratyphi A isolates and negative to 20 other Salmonella and 15 non-Salmonella bacteria. This assay detected 8 samples as positive and 52 samples as negative for S. Paratyphi A. These results were similar to those results obtained by PCR assay and culture methods. Conclusion An in-house LAMP method was established in this study and could potentially be used as a rapid, sensitive, specific and cost-effective detection method for S. Paratyphi A especially in low-resource settings. However this LAMP method is only recommended for screening purposes and need to be further confirmed with gold standard method which is culture method and PCR. |
| Databáze: | OpenAIRE |
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