Abstract
Septicemic melioidosis is the most severe form of melioidosis, which is caused by Burkholderia pseudomallei. It is endemic in Southeast Asia and is the leading cause of death from community-acquired septicemia in northeast Thailand. A major factor that contributes to the high mortality is the delay in isolation and identification of the causative organism. More than half of the patients die within the first 2 days after hospital admission, before bacterial cultures become positive. The present study was undertaken to develop a rapid diagnostic method for identification of this organism. A nested PCR system that amplified a part of 16S rRNA gene that was highly specific to B. pseudomallei was developed. This system was able to detect as few as two bacteria present in the PCR. DNAs from all 30 clinical isolates of B. pseudomallei and none of the other bacteria tested were amplified. The described PCR system has been employed for the detection of the organism in clinical specimens, including buffy coat and pus from internal organs. The detection of B. pseudomallei in buffy coat specimens by PCR was shown to be comparable to the detection of bacteria from blood cultures in septicemic melioidosis cases.
Full Text
The Full Text of this article is available as a PDF (387.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson B. Broad-range polymerase chain reaction for detection and identification of bacteria. J Fla Med Assoc. 1994 Dec;81(12):835–837. [PubMed] [Google Scholar]
- Anuntagool N., Rugdech P., Sirisinha S. Identification of specific antigens of Pseudomonas pseudomallei and evaluation of their efficacies for diagnosis of melioidosis. J Clin Microbiol. 1993 May;31(5):1232–1236. doi: 10.1128/jcm.31.5.1232-1236.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chaowagul W., White N. J., Dance D. A., Wattanagoon Y., Naigowit P., Davis T. M., Looareesuwan S., Pitakwatchara N. Melioidosis: a major cause of community-acquired septicemia in northeastern Thailand. J Infect Dis. 1989 May;159(5):890–899. doi: 10.1093/infdis/159.5.890. [DOI] [PubMed] [Google Scholar]
- Dance D. A. Melioidosis: the tip of the iceberg? Clin Microbiol Rev. 1991 Jan;4(1):52–60. doi: 10.1128/cmr.4.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Desakorn V., Smith M. D., Wuthiekanun V., Dance D. A., Aucken H., Suntharasamai P., Rajchanuwong A., White N. J. Detection of Pseudomonas pseudomallei antigen in urine for the diagnosis of melioidosis. Am J Trop Med Hyg. 1994 Nov;51(5):627–633. doi: 10.4269/ajtmh.1994.51.627. [DOI] [PubMed] [Google Scholar]
- Kitchin P. A., Szotyori Z., Fromholc C., Almond N. Avoidance of PCR false positives [corrected]. Nature. 1990 Mar 15;344(6263):201–201. doi: 10.1038/344201a0. [DOI] [PubMed] [Google Scholar]
- Leelarasamee A., Bovornkitti S. Melioidosis: review and update. Rev Infect Dis. 1989 May-Jun;11(3):413–425. doi: 10.1093/clinids/11.3.413. [DOI] [PubMed] [Google Scholar]
- Lew A. E., Desmarchelier P. M. Detection of Pseudomonas pseudomallei by PCR and hybridization. J Clin Microbiol. 1994 May;32(5):1326–1332. doi: 10.1128/jcm.32.5.1326-1332.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- White N. J., Dance D. A., Chaowagul W., Wattanagoon Y., Wuthiekanun V., Pitakwatchara N. Halving of mortality of severe melioidosis by ceftazidime. Lancet. 1989 Sep 23;2(8665):697–701. doi: 10.1016/s0140-6736(89)90768-x. [DOI] [PubMed] [Google Scholar]
- Yabuuchi E., Kosako Y., Oyaizu H., Yano I., Hotta H., Hashimoto Y., Ezaki T., Arakawa M. Proposal of Burkholderia gen. nov. and transfer of seven species of the genus Pseudomonas homology group II to the new genus, with the type species Burkholderia cepacia (Palleroni and Holmes 1981) comb. nov. Microbiol Immunol. 1992;36(12):1251–1275. doi: 10.1111/j.1348-0421.1992.tb02129.x. [DOI] [PubMed] [Google Scholar]