Comparison of different PCR approaches for characterization of Burkholderia (Pseudomonas) cepacia isolates.
نویسندگان
چکیده
In this study, we evaluated three PCR methods for epidemiological typing of Burkholderia (Pseudomonas) cepacia--PCR-ribotyping, arbitrarily primed PCR (AP-PCR) and enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR)--and compared them with pulsed-field gel electrophoresis. The analysis was performed with 31 isolates of B. cepacia, comprising 23 epidemiologically unrelated isolates and 8 isolates collected from the same patient during two episodes of bacteremia. Pulsed-field gel electrophoresis, ERIC-PCR, and AP-PCR identified 23 distinct types among the 23 unrelated isolates, while PCR-ribotyping only identified 12 strain types, even after AluI digestion of the amplification products. Among the eight isolates collected from the same patient, all typing techniques revealed two clones of strains. The day-to-day reproducibilities of PCR-ribotyping and ERIC-PCR were good, while greater day-to-day variations were noted in the fingerprints obtained by AP-PCR. We conclude that all three PCR techniques are useful for rapid epidemiological typing of B. cepacia, but ERIC-PCR seems to be more reproducible and discriminative.
منابع مشابه
Characterization of PCR-ribotyping for Burkholderia (Pseudomonas) cepacia.
Ribotyping, a method of genotyping bacterial isolates for epidemiologic study, uses rRNA as a probe to detect chromosomal restriction fragment length polymorphisms. Although ribotyping is accurate, its utility is limited by the labor and time necessary for Southern blot analysis. PCR-ribotyping uses PCR to amplify the 16S-23S intergenic spacer region of the bacterial rRNA operon. Length heterog...
متن کاملIsolation and Characterization of Burkholderia Cepacia Strains from Hospitalized Patients in the Hospitals of West Guilan Province
Abstract Background and Objective: Burkholderia cepacia complex (BCC) is a plant pathogen that is an important mortality factor in immune-compromised and hospitalized patients. We aimed to Isolate and Characterize the Burkholderia Cepacia Strains from Hospitalized Patients in the Hospitals of West Guilan Province. Material and Methods: This study was conducted on 90 saliva and blood ...
متن کاملInterspecies biofilms of Pseudomonas aeruginosa and Burkholderia cepacia.
The leading cause of morbidity and mortality in cystic fibrosis (CF) continues to be lung infections with Pseudomonas aeruginosa biofilms. Co-colonization of the lungs with P aeruginosa and Burkholderia cepacia can result in more severe pulmonary disease than P. aeruginosa alone. The interactions between P. aeruginosa biofilms and B. cepacia are not yet understood; one possible association bein...
متن کاملIdentification of isolates within the Burkholderia cepacia complex by a multiplex recA and 16S rRNA gene real-time PCR assay.
Although identification of members of the Burkholderia cepacia complex (BCC) can be accomplished with phenotypic methods (5), this tends to remain the domain of reference laboratories. However, the genetic sequence of the recA gene has also been found to be useful for BCC genomovar identification (6, 8). We describe the use of a monochrome (SYBR green I), real-time PCR assay with simultaneous d...
متن کاملDevelopment of a PCR probe test for identifying Pseudomonas aeruginosa and Pseudomonas (Burkholderia) cepacia.
AIMS To develop a system of species specific polymerase chain reaction (PCR) and DNA hybridisation based on 16s ribosomal RNA sequences for the identification of Pseudomonas aeruginosa and Pseudomonas (Burkholderia) cepacia in sputum from children with cystic fibrosis. METHODS Most of the 16s rRNA sequences from strains of Ps aeruginosa, Ps (Burkholderia) cepacia, and Ps putida were determine...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of clinical microbiology
دوره 33 12 شماره
صفحات -
تاریخ انتشار 1995