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Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction.
Mol Cell Probes. 1997 Feb; 11(1):25-31.MC

Abstract

A highly sensitive, specific, rapid and simple method to detect Burkholderia pseudomallei in blood samples was developed. Two 22-base oligonucleotide primers, based on sequences from a specific DNA probe, were used for amplification of bacterial DNA by the polymerase chain reaction (PCR). Amplification with these primers yielded a 178-base pair product in 100 clinical isolates of B. pseudomallei. As little as 0.5 fg of B. pseudomallei DNA was detectable by this method. Experiments involving inoculation of the organism into uninfected blood samples showed that the method could be used to detect as few as 1 bacterial cell ml-1 of whole blood. Non-specific amplification of other bacterial DNAs from 18 samples of bacteria was not observed. Blood samples from seven patients proven to have melioidosis by haemoculture were positive using these primers. The total time required for sample processing, amplification and visualization was approximately 3.5 h. The high sensitivity, rapidity and simplicity of this method should make it valuable for diagnosis, monitoring of drug treatment and for epidemiological studies of the melioidosis.

Authors+Show Affiliations

Department of Oral Biology, Faculty of Dentistry, Khon Kaen University, Thailand.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9076711

Citation

Rattanathongkom, A, et al. "Detection of Burkholderia Pseudomallei in Blood Samples Using Polymerase Chain Reaction." Molecular and Cellular Probes, vol. 11, no. 1, 1997, pp. 25-31.
Rattanathongkom A, Sermswan RW, Wongratanacheewin S. Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction. Mol Cell Probes. 1997;11(1):25-31.
Rattanathongkom, A., Sermswan, R. W., & Wongratanacheewin, S. (1997). Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction. Molecular and Cellular Probes, 11(1), 25-31.
Rattanathongkom A, Sermswan RW, Wongratanacheewin S. Detection of Burkholderia Pseudomallei in Blood Samples Using Polymerase Chain Reaction. Mol Cell Probes. 1997;11(1):25-31. PubMed PMID: 9076711.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of Burkholderia pseudomallei in blood samples using polymerase chain reaction. AU - Rattanathongkom,A, AU - Sermswan,R W, AU - Wongratanacheewin,S, PY - 1997/2/1/pubmed PY - 1997/2/1/medline PY - 1997/2/1/entrez SP - 25 EP - 31 JF - Molecular and cellular probes JO - Mol Cell Probes VL - 11 IS - 1 N2 - A highly sensitive, specific, rapid and simple method to detect Burkholderia pseudomallei in blood samples was developed. Two 22-base oligonucleotide primers, based on sequences from a specific DNA probe, were used for amplification of bacterial DNA by the polymerase chain reaction (PCR). Amplification with these primers yielded a 178-base pair product in 100 clinical isolates of B. pseudomallei. As little as 0.5 fg of B. pseudomallei DNA was detectable by this method. Experiments involving inoculation of the organism into uninfected blood samples showed that the method could be used to detect as few as 1 bacterial cell ml-1 of whole blood. Non-specific amplification of other bacterial DNAs from 18 samples of bacteria was not observed. Blood samples from seven patients proven to have melioidosis by haemoculture were positive using these primers. The total time required for sample processing, amplification and visualization was approximately 3.5 h. The high sensitivity, rapidity and simplicity of this method should make it valuable for diagnosis, monitoring of drug treatment and for epidemiological studies of the melioidosis. SN - 0890-8508 UR - https://www.unboundmedicine.com/medline/citation/9076711/Detection_of_Burkholderia_pseudomallei_in_blood_samples_using_polymerase_chain_reaction_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0890-8508(96)90072-9 DB - PRIME DP - Unbound Medicine ER -