Evaluation of microbial RNA extractions from Streptococcus pneumoniae.

Evaluation of microbial RNA extractions from Streptococcus pneumoniae. Research Abstract Details 

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Evaluation of microbial RNA extractions from Streptococcus pneumoniae. Abstract Text:

Ha-Sheng Li-Korotky,Lori A Kelly,Otavio Piltcher,Patricia A Hebda,William J Doyle,

The mechanisms that control Streptococcus pneumoniae's ability to colonize the nasopharynx or to invade the middle ear and cause acute otitis media are not understood. Focused study of these mechanisms requires efficient methods for the extraction of microbial RNA from minute clinical samples. Several lysis/extraction methods were tested and compared to determine the optimal conditions for isolating intact total RNA from pneumococcal cells. The sensitivity and efficiency of the extractions were evaluated by reverse transcription polymerase chain reaction (RT-PCR). Compared to other methods, mechanical homogenization in TRIZOL was the most efficient for releasing microbial RNA, and addition of polyinosinic acid (Poly I) as an RNA carrier increased the assay sensitivity to 10(2) colony forming units when detected by RT-PCR amplification of 16S ribosomal RNA or messenger RNA for penicillin binding protein 2b. Quantitative results were confirmed using a ribonuclease protection assay. Penicillin binding protein 2b was also detected in rat middle ear mucosa recovered 5 weeks after middle ear challenge with S. pneumoniae. This study describes a useful core methodology for use in identifying pneumococcal virulence genes from small titer samples and has promising applications in clinical studies of pneumococcal nasopharyngeal colonization and otitis media pathogenesis.

Evaluation of microbial RNA extractions from Streptococcus pneumoniae. Publishing Authors By Initials

HS Li-Korotky,LA Kelly,O Piltcher,PA Hebda,WJ Doyle,

For similar bacteria: gram-positive bacteria: gram-positive cocci: streptococcaceae: streptococcus: streptococcus pneumoniae research abstracts see: bacteria: gram-positive bacteria: gram-positive cocci: streptococcaceae: streptococcus: streptococcus pneumoniae research

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Evaluation of microbial RNA extractions from Streptococcus pneumoniae. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Journal of microbiological methods

VOLUME: 68

Page Numbers: 342-8

Journal Abbreviation: J. Microbiol. Methods

ISSN: 0167-7012

DAY: 13

MONTH: 11

YEAR: 2006

Evaluation of microbial RNA extractions from Streptococcus pneumoniae. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 8306883

Evaluation of microbial RNA extractions from Streptococcus pneumoniae. Keywords Mesh Terms:

KEYWORDS: Streptococcus pneumoniae

MESH TERMS: isolation & purification

Chemical & Substance for Abstract: Evaluation of microbial RNA extractions from Streptococcus pneumoniae. Information

Substance Name: mutanolysin

Registry Number: EC 3.4.99.-

Grant and Affiliation Information for Evaluation of microbial RNA extractions from Streptococcus pneumoniae.

AFFILIATION: Division of Pediatric Otolaryngology, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania, 15213, USA. Ha-Sheng.Li@chp.edu

Country: Netherlands

AGENCY: United States NIDCD

GRANT: R03 DC007511-02

ACRONYM: DC

MEDLINETA: J Microbiol Methods

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