scholarly journals Rapid Molecular Diagnosis of Lactobacillus Bacteremia by Terminal Restriction Fragment Length Polymorphism Analysis of the 16S rRNA Gene

2004 ◽  
Vol 2 (1) ◽  
pp. 37-45 ◽  
Author(s):  
J. E. Christensen ◽  
C. E. Reynolds ◽  
S. K. Shukla ◽  
K. D. Reed
2003 ◽  
Vol 69 (2) ◽  
pp. 1251-1262 ◽  
Author(s):  
Koji Nagashima ◽  
Takayoshi Hisada ◽  
Maremi Sato ◽  
Jun Mochizuki

ABSTRACT New primer-enzyme combinations for terminal restriction fragment length polymorphism (T-RFLP) targeting of the 16S rRNA gene were constructed by using the T-RFLP analysis program (designated TAP T-RFLP) located at the Ribosomal Database Project website, and their performance was examined empirically. By using the fluorescently labeled 516f primer (Escherichia coli positions 516 to 532) and 1510r primer (positions 1510 to 1492), the 16S rRNA gene was amplified from human fecal DNA. The resulting amplified product was digested with RsaI plus BfaI or with BslI. When the T-RFLP was carried out with fecal DNAs from eight individuals, eight predominant operational taxonomic units (OTUs) were detected with RsaI and BfaI digestion and 14 predominant OTUs were detected with BslI digestion. The distribution of the OTUs was consistent with the results of the computer simulations with TAP T-RFLP. The T-RFLP analyses of the fecal DNAs from individuals gave characteristic profiles, while the variability of the T-RFLP profiles between duplicate DNA preparations from the same samples were minimal. This new T-RFLP method made it easy to predict what kind of intestinal bacterial group corresponded to each OTU on the basis of the terminal restriction fragment length compared with the conventional T-RFLP and, moreover, made it possible to identify the bacterial species that an OTU represents by cloning and sequencing.


1999 ◽  
Vol 37 (12) ◽  
pp. 4158-4160 ◽  
Author(s):  
Stephen M. Marshall ◽  
Pasquale L. Melito ◽  
David L. Woodward ◽  
Wendy M. Johnson ◽  
Frank G. Rodgers ◽  
...  

A rapid two-step identification scheme based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the 16S rRNA gene was developed in order to differentiate isolates belonging to theCampylobacter, Arcobacter, andHelicobacter genera. For 158 isolates (26 reference cultures and 132 clinical isolates), specific RFLP patterns were obtained and species were successfully identified by this assay.


2006 ◽  
Vol 72 (5) ◽  
pp. 3788-3792 ◽  
Author(s):  
Julie C. Frey ◽  
Jessica M. Rothman ◽  
Alice N. Pell ◽  
John Bosco Nizeyi ◽  
Michael R. Cranfield ◽  
...  

ABSTRACT We describe the bacterial diversity in fecal samples of a wild gorilla by use of a 16S rRNA gene clone library and terminal-restriction fragment length polymorphism (T-RFLP). Clones were classified as Firmicutes, Verrucomicrobia, Actinobacteria, Lentisphaerae, Bacteroidetes, Spirochetes, and Planctomycetes. Our data suggest that fecal populations did not change temporally, as determined by T-RFLP.


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