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PCR-RFLP based differentiation of Burkholderia mallei and Burkholderia pseudomallei.
Mol Cell Probes. 2004 Apr; 18(2):97-101.MC

Abstract

Burkholderia mallei and Burkholderia pseudomallei manifest a high similarity with regard to clinical syndromes, glanders and melioidosis. Phenotypic and genotypic characters are also highly similar. In an attempt to differentiate the two organisms, the molecular method was applied. This study aimed to identify the different DNA fragment in B. mallei, as compared with B. pseudomallei. The Sau3AI-digested genomic DNA patterns of B. mallei and B. pseudomallei are distinctive, especially the DNA fragments between 0.9-1.5 Kb in size. A 900-bp specific DNA fragment of B. mallei was cloned and sequenced. Using the specific DNA fragment as a probe, Southern blot hybridization was performed to differentiate the two species. The results of hybridization patterns are effective in to elucidating the genetic dissimilarities among these two Burkholderia species. Furthermore, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) digested with Sau3AI was developed to allow a more reliable and rapid identification of the two species. A 650-bp PCR-RFLP product of B. mallei was detected, while two fragments of 250 and 400-bp PCR-RFLP products of B. pseudomallei were visualized. The results suggest that the specific DNA fragment in our study should be of considerable use as a genetic marker for ensuring identification of the two species.

Authors+Show Affiliations

Department of Biochemistry, Faculty of Science, Mahidol University, RamaVI Road, Bangkok 10400, Thailand.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

15051118

Citation

Tanpiboonsak, Suda, et al. "PCR-RFLP Based Differentiation of Burkholderia Mallei and Burkholderia Pseudomallei." Molecular and Cellular Probes, vol. 18, no. 2, 2004, pp. 97-101.
Tanpiboonsak S, Paemanee A, Bunyarataphan S, et al. PCR-RFLP based differentiation of Burkholderia mallei and Burkholderia pseudomallei. Mol Cell Probes. 2004;18(2):97-101.
Tanpiboonsak, S., Paemanee, A., Bunyarataphan, S., & Tungpradabkul, S. (2004). PCR-RFLP based differentiation of Burkholderia mallei and Burkholderia pseudomallei. Molecular and Cellular Probes, 18(2), 97-101.
Tanpiboonsak S, et al. PCR-RFLP Based Differentiation of Burkholderia Mallei and Burkholderia Pseudomallei. Mol Cell Probes. 2004;18(2):97-101. PubMed PMID: 15051118.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - PCR-RFLP based differentiation of Burkholderia mallei and Burkholderia pseudomallei. AU - Tanpiboonsak,Suda, AU - Paemanee,Atchara, AU - Bunyarataphan,Sasinee, AU - Tungpradabkul,Sumalee, PY - 2003/07/10/received PY - 2003/09/26/accepted PY - 2004/3/31/pubmed PY - 2004/10/22/medline PY - 2004/3/31/entrez SP - 97 EP - 101 JF - Molecular and cellular probes JO - Mol Cell Probes VL - 18 IS - 2 N2 - Burkholderia mallei and Burkholderia pseudomallei manifest a high similarity with regard to clinical syndromes, glanders and melioidosis. Phenotypic and genotypic characters are also highly similar. In an attempt to differentiate the two organisms, the molecular method was applied. This study aimed to identify the different DNA fragment in B. mallei, as compared with B. pseudomallei. The Sau3AI-digested genomic DNA patterns of B. mallei and B. pseudomallei are distinctive, especially the DNA fragments between 0.9-1.5 Kb in size. A 900-bp specific DNA fragment of B. mallei was cloned and sequenced. Using the specific DNA fragment as a probe, Southern blot hybridization was performed to differentiate the two species. The results of hybridization patterns are effective in to elucidating the genetic dissimilarities among these two Burkholderia species. Furthermore, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) digested with Sau3AI was developed to allow a more reliable and rapid identification of the two species. A 650-bp PCR-RFLP product of B. mallei was detected, while two fragments of 250 and 400-bp PCR-RFLP products of B. pseudomallei were visualized. The results suggest that the specific DNA fragment in our study should be of considerable use as a genetic marker for ensuring identification of the two species. SN - 0890-8508 UR - https://www.unboundmedicine.com/medline/citation/15051118/PCR_RFLP_based_differentiation_of_Burkholderia_mallei_and_Burkholderia_pseudomallei_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0890850803000951 DB - PRIME DP - Unbound Medicine ER -