Comparison of Rapid, Automated Ribotyping and DNA Macrorestriction Analysis of
            <i>Burkholderia pseudomallei</i>

Inglis, Timothy J. J.; O'Reilly, Lyn C.; Foster, Niki F.; Clair, Adele; Sampson, J.

doi:10.1128/jcm.40.9.3198-3203.2002

Cited by 37 publications

(25 citation statements)

References 16 publications

(15 reference statements)

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“…There was no significant difference between the rest of the groups (p=0.69) (Figure 4). Further analysis revealed that two pairs of strains from the Northern Territory clinical group were indistinguishable, while the remaining 7 were classified as distinct by DNA macro restriction with Xbal (Inglis et al, 2002). There was no significant difference in chlorine tolerance between soil and water environmental isolates from Western Australia; a result that was anticipated since most of these isolates were from a single cluster and had a high homology by PFGE.…”

Section: Survival Of Bacteria In Chlorinementioning

confidence: 89%

The effect of free chlorine on Burkholderia pseudomallei in potable water

Howard¹,

Inglis²

2003

Water Research

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Chlorine is widely used in public water supplies to provide a disinfection barrier. The effect of chlorine disinfection on the water-borne pathogen Burkholderia pseudomallei was assessed using multiple techniques. After exposure to chlorine viable bacteria were undetectable by conventional plate count techniques, however persistence of B. pseudomallei was verified by flow cytometry and bacteria were recoverable following a simple one step broth procedure. The minimum residual chlorine concentration and contact time as prescribed by potable water providers in Australia was insufficient to reduce a B. pseudomallei population by more than 2 log 10 . Chlorine had a bacteriostatic effect only on B. pseudomallei; viable bacteria were recovered from water containing up to 1000 ppm free chlorine. This finding has practical implications for water treatment in regions where B. pseudomallei is endemic. Future work to assess the effect of alternative water disinfection processes either singly or in sequence is necessary.

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Section: Survival Of Bacteria In Chlorinementioning

confidence: 89%

The effect of free chlorine on Burkholderia pseudomallei in potable water

Howard¹,

Inglis²

2003

Water Research

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“…A second, complementary genotyping method was introduced that used an automated ribotyping platform, and the two methods were compared. 8 The addition of genotype analytical software (Bionumerics, Kortrijk, Belgium) allowed for the assembly of a genotype archive based on the clinical and environmental isolates steadily amassing in the WA culture collection. As other PCRbased genotyping methods were introduced, the question arose as to which method should serve as the first-line genotyping system and what should be the definitive typing method.…”

Section: Introductionmentioning

confidence: 99%

The Aftermath of the Western Australian Melioidosis Outbreak

Inglis¹,

O'Reilly²,

Merritt³

et al. 2011

The American Society of Tropical Medicine and Hygiene

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Melioidosis became a notifiable disease in Western Australia (WA) 2 years after the West Kimberley melioidosis outbreak. Two cases of melioidosis caused by the outbreak genotype of Burkholderia pseudomallei (National Collection of Type Cultures [NCTC] 13177) occurred in 1998 and 1999 in persons who visited the outbreak location at the time. No other infections caused by the outbreak strain have been recorded in WA since that time, despite an average of four culture-positive cases per year. Sporadic cases of melioidosis often follow tropical storms and cyclones during summer, and they have been detected outside the endemic area when cyclones travel far inland. In 2007, environmental isolates resembling NCTC 13177 were found 500 km east of the outbreak location after unusually severe weather. Recent whole-genome analysis places NCTC 13177 genetically close to other Australian isolates. Additional biogeographic and ecological studies are needed to establish the relative importance of environmental cofactors in disease pathogenesis.

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“…This protocol enabled assembly of a collection of B. pseudomallei and other Burkholderia species, benchmarked against type culture collection strains and imported B. pseudomallei strains from other research centres. Our Burkholderia Culture Collection has been genotyped by EcoR1 ribotyping and Xba1 DNA macrorestriction analysis 5 . The increasing genetic complexity of the genus Burkholderia made us doubt the wisdom of relying on a single PCR product to confirm the identity of new clinical isolates of B. pseudomallei.…”

Section: Introductionmentioning

confidence: 99%

PCR-based identification of Burkholderia pseudomallei

Merritt

Inglis

Chidlow

et al. 2006

Rev. Inst. Med. trop. S. Paulo

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SUMMARYDNA amplification techniques are being used increasingly in clinical laboratories to confirm the identity of medically important bacteria. A PCR-based identification method has been in use in our centre for 10 years for Burkholderia pseudomallei and was used to confirm the identity of bacteria isolated from cases of melioidosis in Ceará since 2003. This particular method has been used as a reference standard for less discriminatory methods. In this study we evaluated three PCR-based methods of B. pseudomallei identification and used DNA sequencing to resolve discrepancies between PCR-based results and phenotypic identification methods. The established semi-nested PCR protocol for B. pseudomallei 16-23s spacer region produced a consistent negative result for one of our 100 test isolates (BCC #99), but correctly identified all 71 other B. pseudomallei isolates tested. Anomalous sequence variation was detected at the inner, reverse primer binding site for this method. PCR methods were developed for detection of two other B. pseudomallei bacterial metabolic genes. The conventional lpxO PCR protocol had a sensitivity of 0.89 and a specificity of 1.00, while a real-time lpxO protocol performed even better with sensitivity and specificity of 1.00, and 1.00. This method identified all B. pseudomallei isolates including the PCR-negative discrepant isolate. The phaC PCR protocol detected the gene in all B. pseudomallei and all but three B. cepacia isolates, making this method unsuitable for PCR-based identification of B. pseudomallei. This experience with PCR-based B. pseudomallei identification methods indicates that single PCR targets should be used with caution for identification of these bacteria, and need to be interpreted alongside phenotypic and alternative molecular methods such as gene sequencing.

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Comparison of Rapid, Automated Ribotyping and DNA Macrorestriction Analysis of Burkholderia pseudomallei

Cited by 37 publications

References 16 publications

The effect of free chlorine on Burkholderia pseudomallei in potable water

The effect of free chlorine on Burkholderia pseudomallei in potable water

The Aftermath of the Western Australian Melioidosis Outbreak

PCR-based identification of Burkholderia pseudomallei

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