Your browser doesn't support javascript.
loading
Discriminative PCR of Bordetella pertussis from closely related Bordetella species using 16S rDNA Gene
Infection and Chemotherapy ; : 24-31, 2008.
Article in English | WPRIM | ID: wpr-721662
ABSTRACT

BACKGROUND:

Polymerase-chain reaction (PCR) detection is useful to diagnosis of pertussis at initial stage because the growth rate of Bordetella pertussis (B. pertussis) is relatively slow. Currently, the primer set for the insertion sequence IS481 (BP primer) is used widely for PCR detection of B. pertussis. However, the cross-reactivity of BP primer set with Bordetella holmesii (B. holmesii) was reported recently. Therefore, discrimination of B. pertussis and B. holmesii is needed in PCR step. For this reason, we developed new primer sets based on 16S rDNA sequence for diagnostic use and estimated the efficiency of these new primer sets. MATERIALS AND

METHODS:

The specific PCR primers were designed from the aligned sequence matrix of 16S rDNA genes of various Bordetella species. The specificity of designed primers were estimated using clinically important 4 Bordetella species, B. pertussis, B. holmesii, Bordetella parapertussis (B. parapertussis) and Bordetella bronchiseptica (B. bronchiseptica). The sensitivity to B. pertussis of designed primers was also estimated and compared with BP primer set.

RESULTS:

As the results, the developed new primer set successfully distinguished B. pertussis and other Bordetella species containing B. holmesii. In the sensitivity assay, the detectable limits of 16S-F2/16S-R1 primer set for B. pertussis were revealed as 5 pg of genomic DNA and 105 cells/mL of cell suspension. In addition to these, identical results between BP with primer and new primer were obtained in clinical samples.

CONCLUSION:

In this study, the specific primer set for B. pertussis was developed based on 16S rDNA sequence and this primer set did not show cross-reactivity to B. holmesii. In addition to these, the applicability of this primer set to the clinical specimens was also confirmed.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Bordetella / Bordetella pertussis / DNA / DNA, Ribosomal / Whooping Cough / Polymerase Chain Reaction / Sensitivity and Specificity / Bordetella bronchiseptica / Bordetella parapertussis / Discrimination, Psychological Type of study: Diagnostic study / Prognostic study Language: English Journal: Infection and Chemotherapy Year: 2008 Type: Article

Similar

MEDLINE

...
LILACS

LIS

Full text: Available Index: WPRIM (Western Pacific) Main subject: Bordetella / Bordetella pertussis / DNA / DNA, Ribosomal / Whooping Cough / Polymerase Chain Reaction / Sensitivity and Specificity / Bordetella bronchiseptica / Bordetella parapertussis / Discrimination, Psychological Type of study: Diagnostic study / Prognostic study Language: English Journal: Infection and Chemotherapy Year: 2008 Type: Article