Direct Detection of
            <i>Helicobacter Pylori</i>
            in Biopsy Specimens using a High-Throughput Multiple Genetic Detection System

Zhang, Yanmei; Wang, Shiwen; Hu, Binjie; Zhao, Fuju; Xiang, Ping; Ji, Danian; Chen, Fei; Liu, Xiaoli; Feng, Yang; Wu, Yong; Kong, Mimi; Li, Nan; Miao, Yingxin; Jiang, Wen-Rong; Fang, Yi; Zhang, Jinghao; Bao, Zhijun; Olszewski, Michal A.; Hu, Zhao

doi:10.2217/fmb-2016-0149

Cited by 6 publications

(4 citation statements)

References 28 publications

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“…The presence of heteroresistance could also be detected as well as the presence of mixed resistance genotypes. This method was found to be very accurate in comparison to culture and rapid uresase test (RUT) using sequencing when necessary, but a limit was the very poor results of culture obtained in this study (28% inferior to RUT) …”

Section: Molecular Methodsmentioning

confidence: 80%

“…This method was found to be very accurate in comparison to culture and rapid uresase test (RUT) using sequencing when necessary, but a limit was the very poor results of culture obtained in this study (28% inferior to RUT). 32 Attempts are also made to develop an on-chip nucleic acid-based analysis on stools using a device derived from the immiscible filtration assisted by surface tension (IFAST) system to extract DNA, with 5 chambers, followed by a ureC PCR. The added value of this system is to allow DNA purification and preconcentration (×40) within 7 minutes.…”

Section: Molecular Methodsmentioning

confidence: 99%

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Diagnosis of Helicobacter pylori infection

et al. 2017

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Important progress is being made in endoscopic methods which allow clinicians to predict the presence of Helicobacter pylori based on characteristics of gastric mucosa and to obtain targeted biopsies. There are also important developments in molecular methods with various techniques derived from standard PCR, applied both on gastric biopsies and stool specimens. Progress is being made in microfluidic systems to get a reliable diagnosis in a very short time. The interest of the C urea breath test has been confirmed as well as stool antigen tests. Attempts are being made to find biological markers of premalignant conditions by serology, other than pepsinogens.

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Section: Molecular Methodsmentioning

confidence: 80%

Section: Molecular Methodsmentioning

confidence: 99%

Diagnosis of Helicobacter pylori infection

et al. 2017

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“…Multicenter perspective studies enrolling a high number of patients at early phases of gastric diseases and performed with high throughput molecular techniques avoiding time-consuming bacterial culture [96] could improve the knowledge of H. pylori patho-physiology and the management of individuals at risk of GC.…”

Section: Discussionmentioning

confidence: 99%

Differential Helicobacter pylori Plasticity in the Gastric Niche of Subjects at Increased Gastric Cancer Risk

et al. 2019

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Helicobacter pylori (H. pylori) represents an independent risk factor for Gastric Cancer (GC). First Degree Relatives (FDR) of GC subjects and Autoimmune Gastritis (AG) patients are both at increased risk for GC. H. pylori genetic heterogeneity within the gastric niche of FDR and AG individuals has been little explored. To understand whether they exploit an increased H. pylori stability and virulence, 14 AG, 25 FDR, 39 GC and 13 dyspeptic patients (D) were investigated by a cultural PCR-based approach characterizing single colonies-forming-units. We chose three loci within the Cytotoxin-associated gene-A Pathogenicity Island (CagPAI) (cagA,cagE,virB11), vacA, homA and homB as markers of virulence with reported association to GC. Inflammatory/precancerous lesions were staged according to Sydney System. When compared to D, FDR, similarly to GC patients, were associated to higher atrophy (OR = 6.29; 95% CI:1.23–31.96 in FDR; OR = 7.50; 95% CI:1.67–33.72 in GC) and a lower frequency of mixed infections (OR = 0.16; 95% CI:0.03–0.81 in FDR; OR = 0.10; 95% CI:0.02–0.48 in GC). FDR presented also an increased neutrophil infiltration (OR = 7.19; 95% CI:1.16–44.65). Both FDR and GC carried a higher proportion of CagPAI+vacAs1i1mx+homB+ profiles (OR = 2.71; 95% CI: 1.66–4.41 and OR = 3.43; 95% CI: 2.16–5.44, respectively). Conversely, AG patients presented a lower frequency of subtypes carrying a stable CagPAI and vacAs1i1mx. These results underline different H. pylori plasticity in FDR and AG individuals, and thus, a different host-bacterium interaction capacity that should be considered in the context of eradication therapies.

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“…Therefore, combined genetic detection may be applied to obtain a more accurate and complete assessment for individualized treatment of tumors (25). New generation technology in gene detection has enabled the simultaneous detection of multiple genes (26), but the application of gene detection in the diagnosis and treatment of thyroid cancer remains limited, since our understanding about the pathogenesis of thyroid cancer is still insufficient and the molecular markers are limited. Therefore, it is necessary to continuously explore the complex molecular pathways and pathogenesis-related processes of thyroid cancer.…”

Section: Discussionmentioning

confidence: 99%

Generation and identification of a thyroid cancer cell line with stable expression of CCDC67 and luciferase reporter genes

et al. 2019

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Coiled-coil domain containing 67 (CCDC67) gene is a tumor suppressor gene that exhibits a significant inhibitory effect on a variety of tumors. Our previous study demonstrated that the upregulation of CCDC67 gene in TPC-1 cells inhibited cell proliferation, migration and invasion, and promoted apoptosis in vitro. However, due to the lack of a suitable cell tool, these results were not validated in vivo. In the present study, a thyroid cancer cell line with stable expression of CCDC67 and luciferase reporter genes was generated and identified. Firstly, cDNA clones of the CCDC67 gene were obtained by reverse transcription using a custom-designed primer. The results of subsequent electrophoresis analysis and sequencing revealed that the cDNA clones of CCDC67 gene were obtained successfully, with a length of 1,862 bp. The lentiviral vectors, containing the CCDC67, luciferase reporter and puromycin acetyltransferase genes, were co-transfected with two plasmids that encode lentiviral structural proteins and envelope proteins into 293T cells. Following ultracentrifugation, the titer of lentivirus was determined by ELISA to be 5.0×108 TU/ml. The constructed lentiviral vector was used to transfect TPC-1 thyroid cancer cells, and stabilization was achieved by puromycin screening. The expression of CCDC67 gene, luciferase activity and tumorigenic ability of the generated cell line were detected. Reverse transcription-qPCR results demonstrated that the expression levels of CCDC67 gene in TPC-1 cells following transfection were increased 194,46.782-fold compared with those in the negative control group (P<0.01). A higher fluorescence intensity was detected in the generated cell line, while no detectable fluorescence was observed in untransfected TPC-1 cells. The tumorigenic ability of TPC-1-Luc-Puromycin-CCDC67 cells was verified by bioluminescence imaging and histopathological analysis using a pulmonary metastasis model. These results demonstrated that a thyroid cancer cell line with stable expression of CCDC67 and luciferase reporter genes was generated successfully. The TPC-1-Luc-Puromycin-CCDC67 cell line may be a helpful tool for further research on CCDC67 in vivo.

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Direct Detection of Helicobacter Pylori in Biopsy Specimens using a High-Throughput Multiple Genetic Detection System

Cited by 6 publications

References 28 publications

Diagnosis of Helicobacter pylori infection

Diagnosis of Helicobacter pylori infection

Differential Helicobacter pylori Plasticity in the Gastric Niche of Subjects at Increased Gastric Cancer Risk

Generation and identification of a thyroid cancer cell line with stable expression of CCDC67 and luciferase reporter genes

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