One-step polymerase chain reaction-based typing of Helicobacter pylori vacA gene: association with gastric histopathology

Han, Shan-Rui; Schneider, Thomas; Loos, Michael; Bhakdi, Sucharit; Maeurer, Markus

doi:10.1007/s004300050115

Cited by 8 publications

(8 citation statements)

References 37 publications

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“…When both cagA and vacA detection is performed, a strong association exists between the presence of cagA and vacA s1, corresponding to strains with the highest production of cytotoxin (23,202,217,475,547,589) as well as more severe pathology and disease.…”

Section: Molecular Methodsmentioning

confidence: 99%

Helicobacter pyloriDetection and Antimicrobial Susceptibility Testing

2007

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SUMMARY The discovery of Helicobacter pylori in 1982 was the starting point of a revolution concerning the concepts and management of gastroduodenal diseases. It is now well accepted that the most common stomach disease, peptic ulcer disease, is an infectious disease, and all consensus conferences agree that the causative agent, H. pylori, must be treated with antibiotics. Furthermore, the concept emerged that this bacterium could be the trigger of various malignant diseases of the stomach, and it is now a model for chronic bacterial infections causing cancer. Most of the many different techniques involved in diagnosis of H. pylori infection are performed in clinical microbiology laboratories. The aim of this article is to review the current status of these methods and their application, highlighting the important progress which has been made in the past decade. Both invasive and noninvasive techniques will be reviewed.

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Section: Molecular Methodsmentioning

confidence: 99%

Helicobacter pyloriDetection and Antimicrobial Susceptibility Testing

2007

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“…The cagA gene status was determined by the presence or absence of cagA amplicons. The vacA genotypes, including signal(s) sequences and midregion (m) types, were characterized by a one-step PCR method (22). For ribotyping, a 1.5-kb fragment was amplified with broad-specificity 16S ribosomal DNA (rDNA) primer pairs pA-f (5Ј-AGAGTT TGATCCTGGCTCAG-3Ј) and pH-b (5Ј-AAGGAGGTGATCCAGCCGCA-3Ј) (15), using an initial denaturation step at 94°C for 2 min, followed by 35 cycles of denaturation at 94°C, annealing at 55°C, and extension at 72°C (each for 1 min), and a final extension at 72°C for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Helicobacter pylori : Clonal Population Structure and Restricted Transmission within Families Revealed by Molecular Typing

Han¹,

Zschausch²,

Meyer³

et al. 2000

J Clin Microbiol

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Helicobacter pylori infects up to 50% of the human population worldwide. The infection occurs predominantly in childhood and persists for decades or a lifetime. H. pylori is believed to be transmitted from person to person. However, tremendous genetic diversity has been reported for these bacteria. In order to gain insight into the epidemiological basis of this phenomenon, we performed molecular typing of H. pylori isolates from different families. Fifty-nine H. pylori isolates from 27 members of nine families were characterized by using restriction fragment length polymorphism analysis of five PCR-amplified genes, by pulsed-field gel electrophoresis (PFGE) of chromosomal DNA, and byvacA and cagA genotyping. The 16S rRNA gene exhibited little allelic variation, as expected for a unique bacterial species. In contrast, the vacA, flaA,ureAB, and lspA-glmM genes were highly polymorphic, with a mean genetic diversity of 0.83, which exceeds the levels recorded for all other bacterial species. In conjunction with PFGE, 59 H. pylori isolates could be differentiated into 21 clonal types. Each individual harbored only one clone, occasionally with a clonal variant. Identical strains were always found either between siblings or between a mother and her children. Statistical analysis revealed clonality of population structure in all isolates. The results of this study suggest the possible coexistence of a large array of clonal lineages that are evolving in each individual in isolation from one another. Transmission appears to occur primarily from mother to child and perhaps between siblings.

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“…1999 ). One other single‐step vacA genotyping assay has been described previously that used the Atherton primers to amplify the signal region but novel primers to target the mid‐region ( Han et al . 1999 ).…”

Section: Discussionmentioning

confidence: 99%

“…Han et al . (1999 ) reported that their multiplex assay was unable to determine vacA type in 7% of isolates tested.…”

Section: Discussionmentioning

confidence: 99%

Determination of Helicobacter pylori vacA allelic types by single-step multiplex PCR

Chisholm

Teare²,

Patel

et al. 2002

Lett Appl Microbiol

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Aims: To develop and evaluate a novel multiplex PCR assay that enables definition of Helicobacter pylori vacA allelic type in a single reaction. Methods and Results: Application of the one-step system to DNA extracts from 22 cultures of known vacA genotype demonstrated that it was highly accurate. Analysis of 15 matched gastric biopsy/culture pairs generated exactly correlating genotype profiles. vacA genotypes were determined from an additional 62/70 gastric biopsies from dyspeptic patients of known H. pylori positive status by the one-step assay, compared with 63/70 by the original tworeaction test. Types s1/m1, s1/m2 and s2/m2 were identified in 51AE9%, 31AE2% and 16AE9% of biopsies, respectively. Conclusions: The multiplex PCR system developed enables rapid one-step vacA genotyping that is accurate, easy to interpret and more economical than the alternative multiple-reaction tests. Application of this system to gastric biopsies from patients in South-east England demonstrated that s1/m1 was the most common genotype, while s1/m2 and s2/m2 were less prevalent. Significance and Impact of the Study: This simple one-step system can be applied direct to antral gastric biopsies without the need for culture, thereby facilitating rapid surveillance of vacA genotype in relation to geographical location and disease status.

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One-step polymerase chain reaction-based typing of Helicobacter pylori vacA gene: association with gastric histopathology

Cited by 8 publications

References 37 publications

Helicobacter pyloriDetection and Antimicrobial Susceptibility Testing

Helicobacter pyloriDetection and Antimicrobial Susceptibility Testing

Helicobacter pylori : Clonal Population Structure and Restricted Transmission within Families Revealed by Molecular Typing

Determination of Helicobacter pylori vacA allelic types by single-step multiplex PCR

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