Helicobacter pylori pathogenesis and disease outcomes are mediated by a complex interplay between bacterial virulence factors, host, and environmental factors. After H. pylori enters the host stomach, four steps are critical for bacteria to establish successful colonization, persistent infection, and disease pathogenesis: (1) Survival in the acidic stomach; (2) movement toward epithelium cells by flagella-mediated motility; (3) attachment to host cells by adhesins/receptors interaction; (4) causing tissue damage by toxin release. Over the past 20 years, the understanding of H. pylori pathogenesis has been improved by studies focusing on the host and bacterial factors through epidemiology researches and molecular mechanism investigations. These include studies identifying the roles of novel virulence factors and their association with different disease outcomes, especially the bacterial adhesins, cag pathogenicity island, and vacuolating cytotoxin. Recently, the development of large-scale screening methods, including proteomic, and transcriptomic tools, has been used to determine the complex gene regulatory networks in H. pylori. In addition, a more available complete genomic database of H. pylori strains isolated from patients with different gastrointestinal diseases worldwide is helpful to characterize this bacterium. This review highlights the key findings of H. pylori virulence factors reported over the past 20 years.

This study reports a new diagnostic assay for the rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) by combing nucleic acid extraction and isothermal amplification of target nucleic acids in a magnetic bead-based microfluidic system. By using specific probe-conjugated magnetic beads, the target deoxyribonucleic acid (DNA) of the MRSA can be specifically recognized and hybridized onto the surface of the magnetic beads which are then mixed with clinical sample lysates. This is followed by purifying and concentrating the target DNA from the clinical sample lysates by applying a magnetic field. Nucleic acid amplification of the target genes can then be performed by the use of a loop-mediated isothermal amplification (LAMP) process via the incorporation of a built-in micro temperature control module, followed by analyzing the optical density (OD) of the LAMP amplicons using a spectrophotometer. Significantly, experimental results show that the limit of detection (LOD) for MRSA in the clinical samples is approximately 10 fg μL(-1) by performing this diagnostic assay in the magnetic bead-based microfluidic system. In addition, the entire diagnostic protocol, from bio-sample pre-treatment to optical detection, can be automatically completed within 60 min. Consequently, this miniature diagnostic assay may become a powerful tool for the rapid purification and detection of MRSA and a potential point-of-care platform for detection of other types of infections.

The prevalence of three plasmid-mediated quinolone resistance determinants, QnrA, QnrB, and QnrS, among 526 nonreplicate clinical isolates of Enterobacter cloacae collected at a Taiwanese university hospital in 2004 was determined by PCR and colony hybridization, and the association of Qnr with the IMP-8 metallo-␤-lactamase was investigated. Eighty-six (16.3%) of all isolates were qnr positive, and the qnrA1-like, qnrB2-like, and qnrS1-like genes were detected alone or in combination in 3 (0.6%), 53 (10.1%), and 34 (6.5%) isolates, respectively. Among 149 putative extended-spectrum-␤-lactamase-producing isolates, 59 (39.6%) isolates, all of which were SHV-12 producers, harbored qnrA (0.7%; 1 isolate), qnrB (28.9%; 43 isolates), or qnrS (12.1%; 18 isolates). Forty-four (78.6%) of 56 IMP-8 producers carried qnrB (58.9%; 33 isolates), qnrS (25.0%; 14 isolates), or both. PCR and sequence analysis revealed that qnrA1 was located in a complex sul1-type integron that contains dhr15, aadA2, qacE⌬1, sul1, orf513, qnrA1, ampR, and qacE⌬1. Conjugation experiments revealed the coexistence of qnrB and bla IMP-8 on the transferred plasmids and the absence of ␤-lactamase content on the transferred qnrS-positive plasmids. The transferred bla IMP-8 -positive plasmids with and without qnrB had very similar restriction patterns, suggesting the horizontal mobility of qnrB. Pulsed-field gel electrophoresis showed six major patterns among the 44 qnr-positive IMP-8-producing isolates. Thus, the extremely high prevalence of qnr among the metallo-␤-lactamase-producing E. cloacae isolates in the hospital may be due mainly to the intrahospital spread of a few clones and the dissemination of plasmids containing both qnrB and bla IMP-8 .Since the first plasmid-mediated quinolone resistance determinant, Qnr (later termed QnrA), was described in a Klebsiella pneumoniae strain from the United States in 1998 (17), three major groups of Qnr determinants, QnrA, QnrB, and QnrS, have been identified in various enterobacterial species (3-14, 17, 20, 22, 33, 34). Qnr determinants may protect DNA gyrase directly from quinolone inhibition, leading to an 8-to 32-fold increase in MICs of quinolones (31). qnrA has been well known for its worldwide distribution and is located in complex sul1-type class 1 integrons (12,16,20,22,28,33). Such integrons consist of duplicate qacE⌬1 and sul1 genes, which surround a putative recombinase gene, orf513 (23). At least five qnrB and two qnrS variants have been described (1, 7-9, 11, 26), among which only qnrB2 has been found in the complex sul1-type integron (7). QnrB and QnrS have been detected in Asia, Europe, and the United States (1,4,8,9,14,24).In Taiwan, qnrS on a plasmid encoding the SHV-2 extendedspectrum ␤-lactamase (ESBL) from a K. pneumoniae strain has been described (4), and the presence of qnrA and qnrB has not been reported. The present study was conducted to determine the prevalence of the three groups of Qnr among Enterobacter cloacae isolates in a Taiwanese teaching hospital. An extraordinary high fr...

To our knowledge, Flavobacterium indologenes has never been reported as a cause of bacteremia in humans. F. indologenes bacteremia was diagnosed in 12 patients at a tertiary referral center in southern Taiwan between 1 January 1992 and 31 December 1994. Six of these patients had ventilator-associated pneumonia, two had primary bacteremia, and one patient each had pyonephrosis, peritonitis, biliary tract infection, and surgical wound infection. Five patients (42%) had malignancies, and three (25%) had multiple burns. Polymicrobial bacteremia was diagnosed in eight patients (67%). Two (17%) of the patients in this study died; both had polymicrobial bacteremia. Antimicrobial susceptibility testing of the blood isolates from the 12 patients showed that > 90% of the isolates were susceptible to piperacillin, cefoperazone, ceftazidime, and minocycline. The chromatograms of esterified fatty acids for the isolates were identical. F. indologenes should be considered an etiologic agent of bloodstream infection, especially in hospitalized patients with severe underlying diseases.

Helicobacter pylori has developed several mechanisms to evade the intracellular killing after phagocytosis. In this study, we reported that some Taiwanese clinical isolated H. pylori can multiply in human monocytic cells, such as THP-1 or U937 cells, but not in murine macrophage Raw264.7 cells. After internalization, there was a 5- to 10-fold increment of re-cultivable H. pylori from the infected THP-1 cells at 12 hrs post infection. The dividing H. pylori was found in a double-layer vesicle, which is characteristic of autophagosome. The formation of autophagosomes is associated with the multiplication of H. pylori in THP-1 cells. Its modulation with rapamycin or 3-MA affects the level of H. pylori replication. Furthermore, the VacA or CagA mutants of H. pylori have lower levels of multiplication in macrophages. We conclude that H. pylori infection induces autophagosome formation, and these autophagic vesicles were adapted for the multiplication of H. pylori in the host.