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RefWorks
Reference Manager
BibTeX
Zotero
EndNote
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Mehrabadi JF, Morsali P, Nejad HR, Imani Fooladi AA, 2012. Detection of toxigenic Vibrio cholerae with new multiplex PCR. J Infect Public Health 5: 263–267.
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Raychoudhuri A, Mukhopadhyay AK, Ramamurthy T, Nandy RK, Takeda Y, Nair GB, 2008. Biotyping of Vibrio cholerae O1: time to redefine the scheme. Indian J Med Res 128: 695–698.
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Nithya R, Ahmed SA, Hoe CH, Gopinath SC, Citartan M, Chinni SV, Lee LP, Rozhdestvensky TS, Tang TH, 2015. Non-protein coding RNA genes as the novel diagnostic markers for the discrimination of Salmonella species using PCR. PLoS One 10: e0118668.
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Utilization of Small RNA Genes to Distinguish Vibrio cholerae Biotypes via Multiplex Polymerase Chain Reaction
Siti Aminah Ahmed
Siti Aminah Ahmed
Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;
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Carsten A. Raabe
Carsten A. Raabe
Institute of Medical Biochemistry, Centre for Molecular Biology of Inflammation (ZMBE), University of Münster, Münster, Germany;
Brandenburg Medical School (Medizinische Hochschule Brandenburg [MHB]), Neuruppin, Germany;
Institute of Experimental Pathology (ZMBE), University of Münster, Münster, Germany;
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Brandenburg Medical School (Medizinische Hochschule Brandenburg [MHB]), Neuruppin, Germany;
Institute of Experimental Pathology (ZMBE), University of Münster, Münster, Germany;
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Hong Leong Cheah
Hong Leong Cheah
Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;
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Chee Hock Hoe
Chee Hock Hoe
Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;
Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kelantan, Malaysia;
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Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kelantan, Malaysia;
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Timofey S. Rozhdestvensky
Timofey S. Rozhdestvensky
Medical Faculty, Transgenic Animal and Genetic Engineering Models (TRAM), University of Münster, Münster, Germany
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Thean Hock Tang
Thean Hock Tang
Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;
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The diarrheal disease “cholera” is caused by Vibrio cholerae, and is primarily confined to endemic regions, mostly in Africa and Asia. It is punctuated by outbreaks and creates severe challenges to public health. The disease-causing strains are most-often members of serogroups O1 and O139. PCR-based methods allow rapid diagnosis of these pathogens, including the identification of their biotypes. However, this necessitates the selection of specific target sequences to differentiate even the closely related biotypes of V. cholerae. Oligonucleotides for selective amplification of small RNA (sRNA) genes that are specific to these V. cholerae subtypes were designed. The resulting multiplex PCR assay was validated using V. cholerae cultures (i.e., 19 V. cholerae and 22 non–V. cholerae isolates) and spiked stool samples. The validation using V. cholerae cultures and spiked stool suspensions revealed detection limits of 10–100 pg DNA per reaction and 1.5 cells/mL suspension, respectively. The multiplex PCR assay that targets sRNA genes for amplification enables the sensitive and specific detection, as well as the differentiation of V. cholerae—O1 classical, O1 El Tor, and O139 biotypes. Most importantly, the assay enables fast and cheaper diagnosis compared with classic culture-based methods.
- Supplemental Materials (PDF 247 KB)
Author Notes
Authors’ addresses: Siti Aminah Ahmed, Hong Leong Cheah, and Thean Hock Tang, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia, E-mails: asiti2000@usm.my, cheahhl814@gmail.com, and tangth@usm.my. Carsten A. Raabe, Institute of Experimental Pathology (ZMBE), University of Münster, Münster, Germany, Brandenburg Medical School (MHB), Neuruppin, Germany, and Institute of Medical Biochemistry, Centre for Molecular Biology of Inflammation (ZMBE), Institute of Medical Biochemistry, University of Münster, Münster, Germany, E-mail: raabec@uni-muenster.de. Chee Hock Hoe, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia, and Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kelantan, Malaysia, E-mail: hcheehock@umk.edu.my. Timofey S. Rozhdestvensky, Medical Faculty, Transgenic Animal and Genetic Engineering Models (TRAM), University of Muenster, Münster, Germany, E-mail: rozhdest@uni-muenster.de.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Ali M, Nelson AR, Lopez AL, Sack DA, 2015. Updated global burden of cholera in endemic countries. PLoS Negl Trop Dis 9: e0003832.
-
2.
World Health Organization, 2016. Cholera annual report 2015. Wkly Epidemiol Rec 91: 433–440.
-
3.
Okpor MD, 2012. Using fuzzy classifier for cholera analysis. Intern J Sci Res 3: 314–317.
-
4.
Nelson EJ, Harris JB, Morris JG Jr., Calderrwood SB, Camilli A, 2009. Cholera transmission: the host, pathogen and bacteriophage dynamic. Nat Rev Microbiol 7: 693–702.
-
5.
Nair GB, 2002. Guidelines for Drinking-Water Quality. Available at: http://www.who.int/water_sanitation_health/dwq/admicrob6.pdf. Accessed March 26, 2018.
-
6.
Islam A et al. 2013. Indigenous Vibrio cholerae strains from a non-endemic region are pathogenic. Open Biol 3: 120181.
-
7.
Faruque SM, Mekalanos JJ, 2012. Phage-bacterial interactions in the evolution of toxigenic Vibrio cholerae. Virulence 3: 556–565.
-
8.
Nair GB, Safa A, Bhuiyan NA, Nusrin S, Murphy D, Nicol C, Valcanis M, Iddings S, Kubuabola I, Vally H, 2006. Isolation of Vibrio cholerae O1 strains similar to pre-seventh pandemic El Tor strains during an outbreak of gastrointestinal disease in an island resort in Fiji. J Med Microbiol 55: 1559–1562.
-
9.
Chin C-S et al. 2011. The origin of the Haitian cholera outbreak strain. N Engl J Med 364: 33–42.
-
10.
Minukhin VV, Kovalenko NI, 2014. Cholera: Methodical Instructions on the Subject Microbiology, Virology and Immunology for the II and III Year English Media Students of Medical and Dentistry Faculties. Kharkiv: Kharkiv National Medical University, 20.
-
11.
Siddique AK, Cash R, 2014. Cholera outbreaks in the classical biotype era. Curr Top Microbiol Immunol 379: 1–16.
-
12.
Mukhopadhyay AK, Takeda Y, Balakrish NG, 2014. Cholera outbreaks in the El Tor biotype era and the impact of the new El Tor variants. Curr Top Microbiol Immunol 379: 17–47.
-
13.
Vicente ACP, 2011. On the emergence of atypical Vibrio cholerae O1 El Tor & cholera epidemic. Indian J Med Res 133: 366–367.
-
14.
Son MS, Megli CJ, Kovacikova G, Qadri F, Taylor RK, 2011. Characterization of Vibrio cholerae O1 El Tor biotype variant clinical isolates from Bangladesh and Haiti, including a molecular genetic analysis of virulence genes. J Clin Microbiol 49: 3739–3749.
-
15.
Kim EJ et al. 2014. Molecular insights into the evolutionary pathway of Vibrio cholerae O1 atypical El Tor variants. PLoS Pathog 10: e1004384.
-
16.
Udden SMN, Zahid MSH, Biswas K, Ahmad QS, Cravioto A, Nair GB, Mekalanos JJ, Faruque SM, 2008. Acquisition of classical CTX prophage from Vibrio cholerae O141 by El Tor strains aided by lytic phages and chitin-induced competence. Proc Natl Acad Sci USA 105: 11951–11956.
-
17.
Keddy KH, Sooka A, Parsons MB, Njanpop-Lafourcade B-M, Fitchet K, Smith AM, 2013. Diagnosis of Vibrio cholerae O1 infection in Africa. J Infect Dis 208: S23–S31.
-
18.
Griffth DC, Kelly-Hope LA, Miller MA, 2006. Review of reported cholera outbreaks worldwide, 1995–2005. Am J Trop Med Hyg 75: 973–977.
-
19.
Loharikar A et al. 2013. A national cholera epidemic with high case fatality rates—Kenya 2009. J Infect Dis 208: S69–S77.
-
20.
Alam A et al. 2010. Diagnostic limitations to accurate diagnosis of cholera. J Clin Microbiol 48: 3918–3922.
-
21.
Greenhill A, Rosewell A, Kas M, Manning L, Latorre L, Siba P, Horwood P, 2012. Improved laboratory capacity is required to respond better to future cholera outbreaks in Papua New Guinea. Western Pac Surveill Response J 3: 30–32.
-
22.
George CM et al. 2014. Evaluation of enrichment method for the detection of Vibrio cholerae O1 using a rapid dipstick test in Bangladesh. Trop Med Int Health 19: 301–307.
-
23.
Ontweka LN et al. 2016. Cholera rapid test with enrichment step has diagnostic performance equivalent to culture. PLoS One 11: e0168257.
-
24.
Yamasaki E, Sakamoto R, Matsumoto T, Morimatsu F, Kurazono T, Hiroi T, Nair GB, Kurazono H, 2013. Development of an immunochromatographic test strip for detection of cholera toxin. Biomed Res Int 2013: 679038.
-
25.
Zhu K, Dietrich R, Didier A, Doyscher D, Märtlbauer E, 2014. Recent developments in antibody-based assays for the detection of bacterial toxins. Toxins 6: 1325–1348.
-
26.
Barzamini B, Moghbeli M, Soleimani NA, 2014. Vibrio cholerae detection in water and wastewater by polymerase chain reaction assay. Int J Enteric Pathog 2: e20997.
-
27.
Kim H-J, Ryu J-O, Lee S-Y, Kim E-S, Kim H-Y, 2015. Multiplex PCR for detection of the Vibrio genus and five pathogenic Vibrio species with primer sets designed using comparative genomics. BMC Microbiol 15: 239.
-
28.
Yamazaki W, Seto K, Taguchi M, Ishibashi M, Inoue K, 2008. Sensitive and rapid detection of cholera toxin-producing Vibrio cholerae using a loop-mediated isothermal amplification. BMC Microbiol 8: 94.
-
29.
Chua AL, Elina HT, Lim BH, Yean CY, Ravichandran M, Lalitha P, 2011. Development of a dry reagent-based triplex PCR for the detection of toxigenic and non-toxigenic Vibrio cholerae. J Med Microbiol 60: 481–485.
-
30.
Mehrabadi JF, Morsali P, Nejad HR, Imani Fooladi AA, 2012. Detection of toxigenic Vibrio cholerae with new multiplex PCR. J Infect Public Health 5: 263–267.
-
31.
Raychoudhuri A, Mukhopadhyay AK, Ramamurthy T, Nandy RK, Takeda Y, Nair GB, 2008. Biotyping of Vibrio cholerae O1: time to redefine the scheme. Indian J Med Res 128: 695–698.
-
32.
Barzelighi HM, Bakhshi B, Boustanshenas M, 2016. Genetic determinants differences between Vibrio cholerae biotypes. Infect Epidemiol Med 2: 26–30.
-
33.
Ahmed SA, Sandai DA, Musa S, Hoe CH, Riadzi M, Lau KL, Tang TH, 2012. Rapid diagnosis of leptospirosis by multiplex PCR. Malays J Med Sci 19: 9–16.
-
34.
Tang TH, Ahmed SA, Musa M, Zainuddin ZF, 2013. Rapid detection of Mycobacterium tuberculosis in clinical samples by multiplex polymerase chain reaction (mPCR). World J Microbiol Biotechnol 29: 2389–2395.
-
35.
Nithya R, Ahmed SA, Hoe CH, Gopinath SC, Citartan M, Chinni SV, Lee LP, Rozhdestvensky TS, Tang TH, 2015. Non-protein coding RNA genes as the novel diagnostic markers for the discrimination of Salmonella species using PCR. PLoS One 10: e0118668.
-
36.
Tokunaga A, Yamaguchi H, Morita M, Arakawa E, Izumiya H, Watanabe H, Osawa R, 2010. Novel PCR-based genotyping method, using genomic variability between repetitive sequences of toxigenic Vibrio cholerae O1 El Tor and O139. Mol Cell Probes 24: 99–103.
-
37.
Li W, Ying X, Lu Q, Chen L, 2012. Predicting sRNAs and their targets in bacteria. Genomics Proteomics Bioinformatics 10: 276–284.
-
38.
Tang TH, Bachellerie JP, Rozhdestvensky T, Bortolin ML, Huber H, Drungowski M, Elge T, Brosius J, Hüttenhofer A, 2002. Identification of 86 candidates for small non-messenger RNAs from the archaeon Archaeoglobus fulgidus. Proc Natl Acad Sci USA 99: 7536–7541.
-
39.
Vogel J, Bartels V, Tang TH, Churakov G, Slagter-Jäger JG, Hüttenhofer A, Wagner EGH, 2003. RNomics in Escherichia coli detects new sRNA species and indicates parallel transcriptional output in bacteria. Nucl Acids Res 31: 6435–6443.
-
40.
Chinni SV, Raabe CA, Zakaria R, Randau G, Hoe CH, Zemann A, Brosius J, Tang TH, Rozhdestvensky TS, 2010. Experimental identification and characterization of 97 novel npcRNA candidates in Salmonella enterica serovar Typhi. Nucl Acids Res 38: 5893–5908.
-
41.
Krӧger C et al. 2012. The transcriptional landscape and small RNAs of Salmonella enterica serovar Typhimurium. Proc Natl Acad Sci USA 109: E1277–E1286.
-
42.
Raabe CA et al. 2010. A global view of the nonprotein-coding transcriptome in Plasmodium falciparum. Nucleic Acids Res 38: 608–617.
-
43.
Chen XS, Rozhdestvensky TS, Collins LJ, Schmitz J, Penny D, 2007. Combined experimental and computational approach to identify non-protein-coding RNAs in the deep-branching eukayote Giardia intestinalis. Nucleic Acids Res 35: 4619–4628.
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