Evaluation of colorimetric detection methods for Shigella, Salmonella, and Vibrio cholerae by loop-mediated isothermal amplification
- Authors: Soli, Kevin , Kas, Monalisa , Maure, Tobias , Umezaki, Masahiro , Morita, Ayako , Siba, Peter , Greenhill, Andrew , Horwood, Paul
- Date: 2013
- Type: Text , Journal article
- Relation: Diagnostic Microbiology and Infectious Disease Vol. 77, no. 4 (2013), p. 321-323
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- Description: We evaluated loop-mediated isothermal amplification end-point detection methods for Salmonella, Shigella, and Vibrio cholerae. Detection sensitivities were comparable to real-time PCR methods. The colorimetric dyes hydroxynaphthol blue and SYBR Green I showed increased sensitivity when compared to visual and automated turbidity readings. End-point colorimetric dyes promise great utility in developing settings.
Improved laboratory capacity is required to respond better future cholera outbreaks in Papua New Guinea
- Authors: Greenhill, Andrew , Rosewell, Alexander , Kas, Monalisa , Manning, Laurens , Latorre, Leomeldo , Siba, Peter , Horwood, Paul
- Date: 2012
- Type: Text , Journal article
- Relation: Western Pacific Surveillance and Response Vol. 3, no. 2 (2012 2012), p. 1-3
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- Description: Cholera was first detected in Papua New Guinea in July 2009, caused by Vibrio cholerae O1 El Tor serotype Ogawa.1 By late 2011, 15 500 cases had been reported throughout lowland Papua New Guinea with a case fatality rate of 3.2%.2 The epidemic has since slowed, with only sporadic cases reported in Western Province and the Autonomous Region of Bougainville (ARB). Accurate and timely diagnosis is a critical element of the public health response to cholera, yet in low-income countries where the burden of cholera is the greatest, diagnostic services are often limited. Here we report on the diagnostic challenges and the logistical factors that impacted on diagnosis during the first reported outbreak of cholera in Papua New Guinea.
Evaluation of a rapid immunochromatographic assay for the detection of rotavirus, norovirus and adenovirus from children hospitalized with acute watery diarrhea
- Authors: Kas, Monalisa , Maure, Tobias , Soli, Kevin , Umezaki, Masahiro , Morita, Ayako , Bebes, Sauli , Jonduo, Marinjho , Larkins, Jo-Ann , Luang-Suarkia, Dagwin , Siba, Peter , Greenhill, Andrew , Horwood, Paul
- Date: 2013
- Type: Text , Journal article
- Relation: Papua and New Guinea Medical Journal Vol. 56, no. 3-4 (2013), p. 141-144
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- Description: We evaluated the IP-Triple I immunochromatographic rapid test for the detection of rotavirus, norovirus and adenovirus using stool samples from children with diarrhoea. The detection of norovirus and adenovirus was poor compared to polymerase chain reaction assays. However, high sensitivity (92%) and specificity (99%) were obtained for the detection of rotavirus.
Detection of enteric viral and bacterial pathogens associated with paediatric diarrhoea in Goroka, Papua New Guinea
- Authors: Soli, Kevin , Maure, Tobias , Kas, Monalisa , Bande, Grace , Bebes, Sauli , Luang-Suarkia, Dagwin , Siba, Peter , Morita, Ayako , Umezaki, Masahiro , Greenhill, Andrew , Horwood, Paul
- Date: 2014
- Type: Text , Journal article
- Relation: International Journal of Infectious Diseases Vol. 27, no. (2014), p. 54-58
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- Description: Objectives: The aim of this study was to investigate the viral and bacterial causes of acute watery diarrhoea in hospitalized children in Papua New Guinea. Methods: A retrospective analysis was conducted on stool samples collected from 199 children (age > 5 years) admitted to the paediatric ward of Goroka General Hospital from August 2009 through November 2010. A large range of viral and bacterial enteric pathogens were targeted using real-time PCR/RT-PCR assays. Results: Young children were much more likely to be admitted with acute gastroenteritis, with 62.8% of patients aged >1 year and 88.4% aged >2 years. An enteric pathogen was detected in 69.8% (n= 138) of patients. The most commonly detected pathogens were Shigella spp (26.6%), rotavirus (25.6%), adenovirus types 40/41 (11.6%), enterotoxigenic Escherichia coli (11.1%), enteropathogenic E. coli (8.5%), norovirus G2 (6.0%), and Campylobacter spp (4.0%). Norovirus G1, sapovirus, and Salmonella spp were also detected, but below our statistical limit of detection. Vibrio cholerae and astrovirus were not detected in any patients. Mixed infections were detected in 22.1% of patients, with Shigella and rotavirus most commonly detected in co-infections with other pathogens. Conclusions: This study demonstrates that Shigella and rotavirus are the major pathogens associated with acute paediatric gastroenteritis in this setting. © 2014 The Authors.
Isolation of Vibrio cholerae and other enteric microbiota from patients with suspected cholera during the 2009 outbreak in Madang Province, Papua New Guinea
- Authors: Kas, Monalisa , Horwood, Paul , Laman, Moses , Manning, Laurens , Atua, Vincent , Siba, Peter , Greenhill, Andrew
- Date: 2013
- Type: Text , Journal article
- Relation: Papua and New Guinea medical journal Vol. 56, no. 3-4 (2013), p. 110-115
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- Description: When cholera was first detected in Papua New Guinea (PNG) in mid-2009, national diagnostic capacity faced many challenges. This was in part due to the non-endemic status of the outbreak, resulting in few local staff experienced in Vibrio cholerae detection and poor access to the required consumables. The PNG Institute of Medical Research conducted culture on specimens from suspected cholera patients in Madang Province, with presumptive V. cholerae isolates sent to Goroka for confirmation. Of 98 samples analysed 15 were culture positive, with V. cholerae detected by polymerase chain reaction (PCR) in an additional 3 samples. Further analyses were conducted to identify other pathogenic bacteria from thiosulphate citrate bile salt sucrose (TCBS) agar. Molecular-based assays detected enteropathogenic (n = 1) and enterotoxigenic (n = 1) strains of Escherichia coli. No other major enteric pathogens were detected. The low detection rate of V. cholerae at the provincial level reflects challenges in the laboratory diagnosis of cholera and in-country challenges in responding to an outbreak of a non-endemic disease, such as lack of in-country diagnostic expertise and available consumables in the early stages. It also suggests that full aetiological investigations are warranted in future outbreaks of acute watery diarrhoea in PNG to fully elucidate the potentially complex aetiology, which could in turn guide diagnostic, treatment and prevention measures.
Wave 2 strains of atypical Vibrio cholerae El Tor caused the 2009-2011 cholera outbreak in Papua New Guinea
- Authors: Greenhill, Andrew , Mutreja, Ankur , Bulach, Dieter , Belousoff, Matthew , Jonduo, Marinjho , Collins, Deirdre , Kas, Monalisa , Wapling, Johanna , Seemann, Torsten , Lafana, Alice , Dougan, Gordon , Brown, Mark , Horwood, Paul
- Date: 2019
- Type: Text , Journal article
- Relation: Microbial genomics Vol. 5, no. 3 (2019), p. 1-5
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- Description: Vibrio cholerae is the causative agent of cholera, a globally important human disease for at least 200 years. In 2009-2011, the first recorded cholera outbreak in Papua New Guinea (PNG) occurred. We conducted genetic and phenotypic characterization of 21 isolates of V. cholerae, with whole-genome sequencing conducted on 2 representative isolates. The PNG outbreak was caused by an atypical El Tor strain harbouring a tandem repeat of the CTX prophage on chromosome II. Whole-genome sequence data, prophage structural analysis and the absence of the SXT integrative conjugative element was indicative that the PNG isolates were most closely related to strains previously isolated in South-East and East Asia with affiliations to global wave 2 strains. This finding suggests that the cholera outbreak in PNG was caused by an exotic (non-endemic) strain of V. cholerae that originated in South-East Asia.