Identification of <i>Bordetella Pertussis </i>in Nasopharyngeal Swabs Using the Polymerase Chain Reaction: Evaluation of Detection Methods

Lichtinghagen, Ralf; Diedrich-Glaubitz, R.; Horsten, Birgit von

doi:10.1515/cclm.1994.32.3.161

Cited by 26 publications

(25 citation statements)

References 14 publications

(13 reference statements)

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“…The use of PCR has made the rapid diagnosis of many infectious diseases possible, and its use in the diagnosis of pertussis infection is rapidly evolving (6,81,229,337,463,520,572,652,653,693,781). Key factors for the successful application of PCR in the diagnosis of infection by Bordetella spp.…”

Section: Specific Diagnosis Of B Pertussis Infectionsmentioning

confidence: 99%

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Molecular Pathogenesis, Epidemiology, and Clinical Manifestations of Respiratory Infections Due toBordetella pertussisand OtherBordetellaSubspecies

2005

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Bordetella respiratory infections are common in people (B. pertussis) and in animals (B. bronchiseptica). During the last two decades, much has been learned about the virulence determinants, pathogenesis, and immunity of Bordetella. Clinically, the full spectrum of disease due to B. pertussis infection is now understood, and infections in adolescents and adults are recognized as the reservoir for cyclic outbreaks of disease. DTaP vaccines, which are less reactogenic than DTP vaccines, are now in general use in many developed countries, and it is expected that the expansion of their use to adolescents and adults will have a significant impact on reducing pertussis and perhaps decrease the circulation of B. pertussis. Future studies should seek to determine the cause of the unique cough which is associated with Bordetella respiratory infections. It is also hoped that data gathered from molecular Bordetella research will lead to a new generation of DTaP vaccines which provide greater efficacy than is provided by today's vaccines

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Section: Specific Diagnosis Of B Pertussis Infectionsmentioning

confidence: 99%

“…REV. (337,463,466,652,781). In one recent study, PCR results were compared with serologic diagnoses, and PCR had a sensitivity of 61% and a specificity of 88% (337).…”

Section: Specific Diagnosis Of B Pertussis Infectionsmentioning

confidence: 99%

Molecular Pathogenesis, Epidemiology, and Clinical Manifestations of Respiratory Infections Due toBordetella pertussisand OtherBordetellaSubspecies

2005

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“…Secondly, falsenegative results are obtained due to any disturbances in the entire assay, which involves all parts from DNA isolation to detection of the amplified DNA (11), such as inhibitory effects during PCR amplification. Another problem is the clinical relevance of any positive result in an assay with such a high sensitivity like a nested polymerase chain reaction.…”

Section: Introductionmentioning

confidence: 99%

A Competitive Polymerase Chain Reaction Assay for Reliable Identification of Bordetella Pertussis in Nasopharyngeal Swabs

Lichtinghagen

Glaubitz²

1995

CCLM

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Summary:In orderte optimize the identification of clinically relevant quantities of Bordetella pertussis in nasopharyngeal swabs, an automated assay introducing competitive polymerase chain reaction was established. A 183 base pair DNA fragment from a repetitive region of the Bordetella pertussis genome was amplified in a polymerase chain reaction. An internal control DNA with nine base substitutions was coamplified in the same reaction. The differentiation between the amplified B. pertussis DNA and the internal control was based on hybridisation against two different probes using Enzymun Test® DNA Detection (Boehringer Mannheim). Nasopharyngeal swabs from serologically positive patients, clinically diagnosed with whooping cough, serologically negative patients after contact with B. pertussis and a negative group were compared. The advantages of competitive PCR are a reduced risk of false-positive and false-negative results and the possibility to differentiate between the different PCR positive groups.

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“…Vertical PAGE was used for PCR product separation, and the amplicons were visualised by silver staining as described previously (Lichtinghagen et al, 1994). Resulting signals were subdivided semiquantitatively in the categories high, intermediate, low and none according to their band intensities.…”

Section: Polyacrylamide (15%) Gel Electrophoresis (Page)mentioning

confidence: 99%

Oncofoetal fibronectin – a tumour-specific marker in detecting minimal residual disease in differentiated thyroid carcinoma

et al. 2005

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Supposedly, thyrocyte-specific transcripts such as thyroglobulin (Tg) and thyroid-stimulating hormone receptor (TSH-R) were proposed to be useful for the diagnosis of circulating tumour cells in patients suffering from differentiated thyroid carcinoma (DTC). However, several research groups reported blood-borne Tg transcripts in healthy individuals. This study determines in particular the origin of Tg mRNA in nucleated blood cells and analyses whether other tumour-associated sequences are absent in leukocytes, but widely expressed in DTC. Therefore, expression analyses for Tg, TSH-R, cytokeratin 19 (CK 19), human telomerase reverse transcriptase (hTERT) and oncofoetal fibronectin (onfFN) were carried out using cDNAs derived from (1) leukocyte fractions, (2) 18 follicular thyroid carcinomas (FTCs) and 48 papillary thyroid carcinomas (PTCs), and (3) leukocytes of two thyrocyte-depleted individuals treated for C-cell carcinoma of the thyroid. Expression of onfFN was additionally analysed by semiquantitative RT -PCR and by quantitative fluorescence-based real-time PCR. Tg and TSH-R expression was demonstrated not only in both athyroid individuals, but in all leukocyte subgroups tested, while hTERT was absent in resting CD4 þ cells and only weakly expressed in the CD8 þ group. CK 19 was notable in each leukocyte population except for resting CD14 þ , as well as for activated and resting CD19 þ cells. All blood cell fractions proved negative for onfFN mRNA, whereas its presence in thyroid carcinoma was 78/98% (FTC/PTC). Threshold cycle values were calculated at: porphobilinogen deaminase (PBGD) ¼ 25.9570.73 (FTC)/24.5575.43 (PTC) (P ¼ 0.2878); onfFN ¼ 25.4873.15 (FTC)/21.4473.44 (PTC) (*P ¼ 0.0001). Finally, onfFN transcripts were detected in blood samples of six out of nine patients with known DTC metastases, demonstrating a reliable assay functionality. We propose that realtime RT -PCR of onfFN mRNA is superior to other markers in monitoring minimal residual disease in DTC with regard to both assay sensitivity and specificity.

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Identification of Bordetella Pertussis in Nasopharyngeal Swabs Using the Polymerase Chain Reaction: Evaluation of Detection Methods

Cited by 26 publications

References 14 publications

Molecular Pathogenesis, Epidemiology, and Clinical Manifestations of Respiratory Infections Due toBordetella pertussisand OtherBordetellaSubspecies

Molecular Pathogenesis, Epidemiology, and Clinical Manifestations of Respiratory Infections Due toBordetella pertussisand OtherBordetellaSubspecies

A Competitive Polymerase Chain Reaction Assay for Reliable Identification of Bordetella Pertussis in Nasopharyngeal Swabs

Oncofoetal fibronectin – a tumour-specific marker in detecting minimal residual disease in differentiated thyroid carcinoma

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