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Introduction: Data about the prevalence of the A2142C, A2142G, and A2143G mutations in 23S rRNA gene is still limited. The aim of this study was to determine the prevalence of these mutations in 23S rRNA gene of H. pylori vietnamese strains. Methodology: One hundred and sixty-nine patients with H. pylori-positive chronic gastritis were examined. H. pylori was detected by rapid urease test and Polymerase chain reaction (PCR). Total DNA was extracted from gastric biopsy specimens. A2142C, A2142G, and A2143G mutations were detected by DNA sequencing and PCR-restriction fragment length polymorphism (PCR-RFLP). Results: A2143G mutation was detected in 36.1% of samples, A2142G mutation in 3.6%, while A2142C mutation was not found in any case. The mixture of wild-type and mutation strains was found in 50% of specimens with A2142G, in 23% of specimens with A2143G mutation. There was no association of 23S rRNA gene point mutations with gender or age. However, an association between the heterogeneity of mutation and age was evidenced, with mean age of the group of pure A2143G higher than the group of wild-type/A2143G mixture, and rate of the wildtype/A2143G mixture higher in patients under 40 years of age. Conclusion: A2143G mutation was prominent, while A2142C mutation was not found in the 23S rRNA gene. PCR-RFLP has revealed a reliable assay allowing a rapid and cost-effective detection of clarithromycin-resistant strains. This is useful in countries as Vietnam with high prevalence of clarithromycin-resistance before choosing optimal therapy for H. pylori eradication.

Introduction: This research aimed to determine the association of the combination of H. pylori infection and TP53 codon 72 polymorphism with non-cardia gastric cancer (GC) in Vietnam. Methodology: A total of 164 patients with non-cardia GC and 164 patients with peptic ulcer disease or functional dyspepsia in controls matched by sex and age were enrolled. H. pylori infection was diagnosed by rapid urease test and polymerase chain reaction (PCR). The cagA gene-positivity and vacA sm subtypes were determined by multiplex PCR. Genotypes of TP53 codon 72 polymorphism were determined by PCR-restriction fragment length polymorphism. Results: The prevalence of H. pylori infection in GC and control group were 61.6% and 55.4%, respectively. The rates of cagA-positive strains in the two H. pylori-positive groups were 80.2% and 71.4%, respectively. There was no statistically significant difference in TP53 codon 72 genotype distribution between GC group (frequencies of Arg/Arg, Arg/Pro and Pro/Pro genotypes were 31.1%, 43.3% and 25.6%, respectively) and controls (29.3%, 52.4% and 18.3%, respectively), p = 0.172. The significant difference in genotype distribution was observed in recessive model (Pro/Pro vs Arg/Arg + Arg/Pro) when stratifying by H. pylori infection (OR = 2.02, 95% CI 1.03–3.96, p = 0.041) and by cagA-positivity (OR = 2.33, 95% CI 1.07–5.07, p = 0.032). Conclusions: This study suggests a synergistic interaction between H. pylori infection, especially cagA-positive H. pylori, and Pro/Pro genotype of TP53 codon 72 polymorphism might play a significant role in the pathogenesis of GC in the Vietnamese population.

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Abstractα-Thalassemia is a common inherited blood disorder manifested mainly by the deletions of α-globin genes. In geographical areas with high carrier frequencies, screening of α-thalassemia carrier state is therefore of vital importance. This study presents a novel method for identifying female carriers of common α-thalassemia deletions using samples routinely taken for non-invasive prenatal tests for screening of fetal chromosomal aneuploidies. A total of 68,885 Vietnamese pregnant women were recruited and α-thalassemia statuses were determined by gap-PCR, revealing 5344 women (7.76%) carried deletions including αα/−−SEA (4.066%), αα/−α3.7 (2.934%), αα/−α4.2 (0.656%), and rare genotypes (0.102%). A two-stage model was built to predict these α-thalassemia deletions from targeted sequencing of the HBA gene cluster on maternal cfDNA. Our method achieved F1-scores of 97.14–99.55% for detecting the three common genotypes and 94.74% for detecting rare genotypes (−α3.7/−α4.2, αα/−−THAI, −α3.7/−−SEA, −α4.2/−−SEA). Additionally, the positive predictive values were 100.00% for αα/αα, 99.29% for αα/−−SEA, 94.87% for αα/−α3.7, and 96.51% for αα/−α4.2; and the negative predictive values were 97.63%, 99.99%, 99.99%, and 100.00%, respectively. As NIPT is increasingly adopted for pregnant women, utilizing cfDNA from NIPT to detect maternal carriers of common α-thalassemia deletions will be cost-effective and expand the benefits of NIPT.

Introduction: Breast cancer is the most common cancer for women worldwide. HER2 expression and gene copy number are important when determining eligibility for adjuvant therapy and/or chemotherapy medications. One challenging issue for breast cancer testing is intratumoral heterogeneity of HER2 gene amplification. Intratumoral heterogeneity can make it difficult to localize target cells of interest. Serial tissue sections used for independent H&E, IHC and FISH stains also increase the difficulty to localize targets due to cellular truncation. We have developed a system to assess both HER2 expression and gene copy number on the same cell. Method: Immunofluorescence (IF) and Fluorescence in situ Hybridization (FISH) were performed on tissue sections from 19 patients with invasive ductal breast carcinoma. Cases were selected based on prior HER2 FISH results (HER2:Chromosome 17 = ratio) representing unamplified (<2.0), amplified (≥2.0) and equivocal (1.8–2.2). Samples were collected from June 2011 – February 2012. Tissue sections were cut at 4uM from formalin-fixed paraffin-embedded tissue blocks. Slides were stained with antibodies for HER2 (Clone #D8F12, Cell Signaling, Danvers, MA), cytokeratin (Clone #AE1, eBioscience, San Diego, CA) and Pan cytokeratin (Clone #PCK-26, Sigma-Aldrich, St. Louis, MO). The whole tissue imaging was performed on the In-Cell (GE Healthcare, Chalfont St. Giles, UK) at 10X. Proprietary software developed by GRC (GE Global Research, Niskayuna, NY) controlled the hardware and performed numerous algorithmic functions. Regions of Interest (ROI) were selected by a pathologist on a whole tissue image and coordinates were recorded by the software. The slides were then imaged at 40x using the previously recorded ROI's. The same slides were stained with the PathVysion HER2/CEP17 FISH kit (Abbott Molecular, Des Plaines, IL). Slides were registered to the previous IF scan using recorded coordinates and tissue morphology recognition algorithms. The sections were imaged for FISH at 40X using the previous ROI selections. Cases were assessed for successful protein and genetic expression using proprietary visualization tools for combined analysis. Results: We evaluated a total of 22 breast cancer cases with 19 cases detecting both protein and gene expression. Of the three cases that could not be evaluated the rationale is as follows: tissue damage incurred during imaging, insufficient focus during the FISH imaging portion, and poor signal to noise of the FISH dots. Conclusion: The reported incidence of intratumoral HER2 amplification heterogeneity is as high as 30%. The challenges associated with tumor heterogeneity may benefit from a standardize analysis method. Using integrated images generated by this system, pathologist is able to select the appropriate cells for HER2 copy number enumeration based on the expression level of HER2 protein, in the same cell, allowing rapid identification of intratumoral heterogeneity. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-05-05.