Helicobacter pylori cagA, vacA, and iceA genotypes and clinical outcomes: a cross-sectional study in central Vietnam.

Helicobacter pylori is the most common cause of gastroduodenal diseases. The concept that cagA-positive H. pylori is a risk factor for gastric cancer appears to be true only for H. pylori strains from Western countries. Other virulent genes may have a synergistic interaction with cagA during pathogenesis. This study aims to investigate H. pylori cagA, vacA, and iceA prevalence, genotypes, and their association to clinical outcomes in Vietnamese patients. The cagA status and vacA and iceA genotypes were determined using the PCR technique on DNA extracted from gastric biopsies of 141 patients with gastroduodenal diseases. After performing molecular analysis for cagA, vacA, and iceA genes, samples with mixed H. pylori strains, positivity, or negativity for both cagA and cagPAI-empty site, or unidentified genotypes were excluded. Finally, 107 samples were examined. The presence of the cagA, vacA, and iceA genes were detected in 77.6%, 100%, and 80.4% of cases, respectively. Notably, cagA( +) with EPIYA-ABD, vacA s1i1m1, vacA s1i1m2, iceA1, and iceA2 accounted for 73.8%, 44.9%, 33.6%, 48.6%, and 31.8% of cases, respectively. Four iceA2 subtypes (24-aa, 59-aa, 94-aa, and 129-aa variants) were found, with the 59-aa variant the most prevalent (70.6%). The cagA( +)/vacAs1i1m1/iceA1 and cagA( +)/vacAs1i1m2/iceA1 combinations were found in 26.2% and 25.1% of cases, respectively. A multivariable logistic regression analysis was performed, after adjusting for age and gender, with the gastritis group was used as a reference control. Statistically significant associations were found between the vacA s1i1m2 genotype, the iceA1 variant, and the cagA( +)/vacAs1i1m2/iceA1 combination and gastric cancer; the adjusted ORs were estimated as 18.02 (95% CI: 3.39-95.81), 4.09 (95% CI: 1.1-15.08), and 16.19 (95% CI: 3.42-76.66), respectively. Interestingly, for the first time, our study found that vacA s1i1m2, but not vacA s1i1m1, was a risk factor for gastric cancer. This study illustrates the genetic diversity of the H. pylori cagA, vacA, and iceA genes across geographical regions and contributes to understanding the importance of these genotypes for clinical outcomes.

Current status of Helicobacter pylori resistance to clarithromycin and levofloxacin in Vietnam: Results from molecular analysis of gastric biopsy specimens.

OBJECTIVES: The management of Helicobacter pylori in Vietnam is becoming progressively more difficult due to increasing antibiotic resistance, particularly to clarithromycin (CLR) and levofloxaxin (LVX). In Vietnam, the selection of an H. pylori eradication regimen is predominantly based on empirical evidence. However, molecular analysis aimed at identifying H. pylori antibiotic-resistant genotypes is a promising method in antibiotic susceptibility testing. In this study, we aimed to determine the rates of genotypic H. pylori resistance to CLR and LVX by using DNA strip technology in Vietnam. METHODS: We performed DNA-strip technology-based testing on 112 patients with H. pylori-positive gastroduodenal diseases to detect 23S rRNA and gyrA mutations. RESULTS: Helicobacter pylori genotypic resistance to CLR and LVX was evident in 81.3% and 53.6% of the patients, respectively, and dual resistance was observed in 48.2%. The 23S rRNA A2142G and A2143G mutations accounted for 1.8% and 79.5% of cases, respectively. The gyrA N87K, D91N, D91G, and D91Y mutations were present in 37.5%, 11.6%, 5.4%, and 5.4% of patients, respectively. All four gyrA mutations were observed in both the naïve and failure patients. We further found an association between the 23S rRNA A2143G mutation and a history of CLR use as well as between the gyrA N87K mutation and a history of LVX use. CONCLUSIONS: We found a very high prevalence of H. pylori resistance to CLR and LVX and dual resistance to these antibiotics in Vietnam. The application of molecular assays is feasible and may improve the management of H. pylori infection in Vietnam.

Genetic diversity of the oipA gene among Helicobacter pylori isolates and clinical outcome in Vietnam.

Outer inflammatory protein A (OipA), which is encoded by the oipA gene, can induce interleukin-8 secretion in gastric epithelial cells. The functional status of the oipA gene is regulated by the slipped-strand mispairing mechanism based on the CT dinucleotide repeat number in the 5' region. This study aimed to investigate the oipA functional status ("on/off") of Helicobacter pylori (H. pylori) and its association with gastroduodenal diseases in southwestern Vietnam. The cross-sectional study was conducted on 173H. pylori isolates from 173 patients with gastroduodenal diseases. Sanger sequencing was used to determine the functional status of oipA. Multivariable logistic regression analysis was performed to identify the association between oipA status and gastroduodenal diseases. The oipA "on" status accounted for 96% of H. pylori isolates. Twenty-five CT repeat patterns of the oipA 5' signal region were observed, five of which were novel CT repeat patterns. The oipA "on" status was found in 100%, 97.8%, and 86.8% of H. pylori isolates from patients with peptic ulcer, precancerous lesions, and chronic gastritis, respectively (p < 0.01). The oipA "on" status was related to gastric precancerous lesions versus chronic gastritis (adjusted OR = 7.39, 95% CI: 1.35-40.59, p = 0.021) and peptic ulcers versus chronic gastritis (adjusted OR = 12.79, 95% CI: 1.19-1760.32, p = 0.033). Our data show a high prevalence of the oipA "on" status, which was associated with precancerous gastric lesions and peptic ulcers. Moreover, genetic diversity in the number and pattern of CT dinucleotide repeat of oipA among Vietnamese H. pylori strains was identified.

Relationship between Allelic Heterozygosity in BoLA-DRB3 and Proviral Loads in Bovine Leukemia Virus-Infected Cattle.

Enzootic bovine leukosis is a lethal neoplastic disease caused by bovine leukemia virus (BLV), belongs to family Retroviridae. The BLV proviral load (PVL) represents the quantity of BLV genome that has integrated into the host's genome in BLV-infected cells. Bovine leukocyte antigen (BoLA) class II allelic polymorphisms are associated with PVLs in BLV-infected cattle. We sought to identify relationships between BoLA-DRB3 allelic heterozygosity and BLV PVLs among different cattle breeds. Blood samples from 598 BLV-infected cattle were quantified to determine their PVLs by real-time polymerase chain reaction. The results were confirmed by a BLV-enzyme-linked immunosorbent assay. Restriction fragment length polymorphism-polymerase chain reaction identified 22 BoLA-DRB3 alleles. Multivariate negative binomial regression modeling was used to test for associations between BLV PVLs and BoLA-DRB3 alleles. BoLA-DRB3.2*3, *7, *8, *11, *22, *24, and *28 alleles were significantly associated with low PVLs. BoLA-DRB3.2*10 was significantly associated with high PVLs. Some heterozygous allele combinations were associated with low PVLs (*3/*28, *7/*8, *8/*11, *10/*11, and *11/*16); others were associated with high PVLs (*1/*41, *10/*16, *10/*41, *16/*27, and *22/*27). Interestingly, the BoLA-DRB3.2*11 heterozygous allele was always strongly and independently associated with low PVLs. This is the first reported evidence of an association between heterozygous allelic combinations and BLV PVLs.

De novo c.2455C>T mutation of NPR2 gene in a fetus with shortened long bones and a ventricular septal defect conceived by a mother with a fragile site at 16q22.1 and a father with a rare heterochromatic variant of chromosome 4 from Vietnam.

BACKGROUND: A heterozygous natriuretic peptide receptor 2 (NPR2) gene c.2455C>T mutation was identified as a cause of familial idiopathic short stature (ISS). Only two cases with this mutation were reported previously, and the probands with ISS had no organ system defects. METHODS: Next-generation sequencing (NGS) was performed on an amniotic fluid DNA sample of a fetus with shortened long bones and a small ventricular septal defect detected by an obstetric ultrasound examination. The pathogenic variant of the fetus was confirmed by Sanger sequencing. Sanger sequencing, G-banded, and C-banded karyotyping of the fetus's parents were subsequently performed. RESULTS: A de novo NPR2 gene c.2455C>T, p.(Arg819Cys) mutation was identified in the fetus. No microdeletion or microduplication was identified in the fetus by copy number variation sequencing with a maximum resolution of 400 kb. The two previous miscarriages experienced by the fetus's parents were interpreted as a result of chromosomal aberrations, including a maternal fragile site at 16q22.1 and a rare paternal variant involving in a large G-band-positive and C-band-positive block of paracentric heterochromatin of chromosome 4p. CONCLUSION: This report provides clinical signs of a de novo heterozygous NPR2 gene c.2455C>T mutation in the fetus and shows paternal chromosomal aberrations causing repeated pregnancy loss.

Establishment of a novel diagnostic test for Bovine leukaemia virus infection using direct filter PCR.

Enzootic bovine leucosis (EBL) is a neoplastic disease of cattle caused by Bovine leukaemia virus (BLV). EBL causes great economic losses, so a fast and reliable diagnostic method is critical for understanding the status of BLV. This will allow us to control BLV infections efficiently and mitigate economic losses. In this study, we established a direct diagnostic test for BLV using dried blood-spotted filter papers without sample pre-treatment. The study was based on 159 clinical blood specimens collected in EDTA from one farm in Kyushu, Japan. The blood-spotted filter papers were used as the template for direct filter PCR. When an ELISA was used as the diagnostic gold standard, the sensitivity and specificity of the direct filter PCR were 90.1% and 97.5%, respectively. The kappa value for the direct filter PCR and real-time PCR methods was 0.97. The dried blood samples spotted onto filter papers were stable for at least 10 days at room temperature, even when the samples were from cattle with a low BLV proviral load. Direct filter PCR is a rapid, easy, reliable and cost-effective diagnostic test that directly detects the BLV proviral genome in clinical blood specimens without DNA extraction. Moreover, it simplifies the collection, transportation and storage procedures for clinical blood specimens.