Feasibility of high resolution melting curve analysis for rapid serotyping of Salmonella from hospitalised patients

Main Article Content

Kritchai Poonchareon
Chaiwat Pulsrikarn
Sukon Khamvichai
Pakpoom Tadee

Abstract

Background: Serum agglutination test is the gold standard phenotyping method widely used for Salmonella enterica characterisation.This conventional method is limited by its complicated and time-consuming procedures. High resolution melting curve (HRM) analysis is introduced as a rapid and labour-saving method.


Objectives: To compare the results of conventional serum agglutination and quantitative PCR-HRM analysis to assess the feasibility of this alternative approach for Salmonella serotyping.


Materials and methods: Salmonella strains from 38 human-originating samples were serotyped using the conventional serum agglutination method and HRM analysis.


Results: The conventional serum agglutination assay detected 14 serotypes, while the HRM analysis identified 10 HRM profiles. There was a correlation between most of the serotyping results obtained by the two methods. Nine of the HRM profiles were unique to a single serotype, of each. One exception was HRM_3. Many of the indistinct curves that were grouped in this HRM pattern belonged to five Salmonella serotypes, including Weltevreden, Corvallis, Derby, Kedougou and Kentucky.


Conclusion: It is difficult to determine all Salmonella serotypes by HRM analysis. However, this method can be used as an alternative to the conventional serum agglutination assay for rapid and labour-saving serotyping.

Article Details

How to Cite
Poonchareon, K., Pulsrikarn, C., Khamvichai, S., & Tadee, P. (2018). Feasibility of high resolution melting curve analysis for rapid serotyping of Salmonella from hospitalised patients. Journal of Associated Medical Sciences, 52(1), 37–41. Retrieved from https://he01.tci-thaijo.org/index.php/bulletinAMS/article/view/134519
Section
Research Articles

References

[1] Antunes P, Mourão J, Pestana N., Peixe L. Leakage of emerging clinically relevant multidrug-resistant Salmonella clones from pig farms. J Antimicrob Chemother 2011; 66(9): 2028-32.

[2] Leader BT, Frye JG, Hu J, Fedorka-Cray PJ, Boyle DS. High-throughput molecular determination of Salmonella enterica serovars by use of multiplex PCR and capillary electrophoresis analysis. J Clin Microbiol 2009; 47(5): 1290-9.

[3] Lopes GV, Michael GB, Cardoso M, Schwarz S. Antimicrobial resistance and class 1 integron-associated gene cassettes in Salmonella enterica
serovar Typhimurium isolated from pigs at slaughter and abattoir environment. Vet Microbiol 2016; 194: 84-92.

[4] Bangtrakulnonth A, Pornreongwong S, Pulsrikarn C, Sawanpanyalert P, Hendriksen RS, Lo Fo Wong DM., et al. Salmonella serovars from humans and other sources in Thailand, 1993-2002. Emerg Infect Dis 2004; 10(1): 131–6.

[5] Popoff MY, Bockemuhl J, McWhorter-Murlin A. Supplement 1991 (no. 35) to the Kauffmann-White scheme. Res Microbiol 1992; 143(8): 807-11.

[6] Sirichote P, Bangtrakulnonth A, Tianmanee K, Unahalekhaka A, Oulai A, Chittaphithakchai P, et al. Serotypes and antimicrobial resistance of Salmonella enterica spp. in central Thailand, 2001-2006. Southeast Asian J Trop Med Public Health 2010; 41(6): 1405-1415.

[7] Herikstad H, Motarjemi Y, Tauxe RV. Salmonella surveillance: a global survey of public health serotyping. Epidemiol Infect 2002; 129(1): 1–8.

[8] Chuanchuen R, Ajariyakhajorn K, Koowatananukul C, Wannaprasat W, Khemtong S, Samngamnim S. Antimicrobial resistance and virulence genes in Salmonella enterica isolates from dairy cows. Food-borne Pathog Dis 2010; 7(1): 63-9.

[9] Foley SL, Lynne AM, Nayak R. Salmonella challenges: prevalence in swine and poultry and potential pathogenicity of such isolates. J Anim Sci 2008; 86 (14 Suppl): E149-62. doi: 10.2527/jas.2007-0464

[10] Grimont PAD, Weill FX. Antigenic formulas of the Salmonella serovar. 9th ed. Paris: Institut Pasteur; 2007.

[11] International Standard Organization. Microbiology of food and animal feeding stuffs - Horizontal method for the detection of Salmonella spp. 4th ed. Geneva: ISO; 2002.

[12] Alvarez J, Sota M, Vivanco AB, Perales I, Cisterna R, Rementeria A, et al. Development of a multiplex PCR method for detection and epidemiological typing of Salmonella in human clinical samples. J Clin Microbiol 2004; 42(4): 1734-8.

[13] Zeinzinger J, Pietzka AT, Stöger A, Kornschober C, Kunert R, Allerberger F. One-step triplex high-resolution melting analysis for rapid identification and simultaneous subtyping of frequently isolated
Salmonella serovars. Appl Environ Microbiol 2012; 78(9): 3352-3360.

[14] Cheng HR, Jiang N. Extremely rapid extraction of DNA from bacteria and yeasts. Biotechnol Lett 2006; 28(1): 55-9

[15] Hunter PR, Gaston MA. Numerical index of the discriminatory ability of typing system: an application of Simpson’s index of diversity. J Clin Microbial 1988; 26(11): 2465-6.

[16] Fowlkes EB, Mallows CL. A method for comparing two hierarchical clusterings. J Am Stat Assoc 1983; 78(383): 553-69.

[17] Bereau of Epidemiology, Thailand. 2017. National Disease Surveillance (Report 506): Food poisoning 2017 [internet]. [Place unknown]: Bereau of Epidemiology; 2017 [cite 2018 April 8]. Available
from: https://www.boe.moph.go.th/boedb/surdata/506wk/y60/d03_5360.pdf.

[18] Huoy L, Pornruangwong S, Pulsrikarn C, Chaturongakul S. Molecular characterization of Thai Salmonella enterica serotype Typhimurium and serotype 4,5,12:i:- reveals distinct genetic deletion pat-terns. Foodborne Pathog Dis 2014; 11(8): 589-92.

[19] Tadee P, Boonkhot P, Pornruangwong S, Patchanee P. Comparative phenotypic and genotypic characterization of Salmonella spp. in pig farms and slaughterhouses in two provinces in northern Thailand. Plos One 2015; 10(2): e0116581. doi: 10.1371/journal.pone.0116581.

[20] Hendriksen RS, Hello SL, Bortolaia V, Pulsrikarn C, Nielsen EM, Pornruangmong S, et al. Characterization of isolates of Salmonella enterica Serovar Stanley, a serovar endemic to asia and associated with travel. J Clin Microbiol 2012; 50(3): 709-20.

[21] Padungtod P, Kaneene JB. Salmonella in food animals and humans in northern Thailand. Int J Micro-biol 2006; 108(3): 346-54.

[22] O’Regan E, McCabe E, Burgess C, McGuinness S, Barry T, Duffy G, et al. Development of a real-time multiplex PCR assay for the detection of multiple Salmonella serotypes in chicken samples. BMC Mi-crobiol. 2002; 8: 156. doi: 10.1186/1471-2180-8-156.

[23] Ren X, Fu Y, Xu C, Feng Z, Li M, Zhang L, et al. High resolution melting (HRM) analysis as a new tool for rapid identification of Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum. Poult Sci 2017; 96(5): 1088-93.