Displaying publications 1 - 20 of 29 in total

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  1. Studies on variation in Lancefield group A type 50 streptococcal cell-wall carbohydrate
    Kurl DN
    Adv Exp Med Biol, 1997;418:607-10.
    PMID: 9331725
    Matched MeSH terms: Antigens, Bacterial/genetics
  2. Detection of virulence genes in Malaysian Shigella species by multiplex PCR assay
    Thong KL, Hoe SL, Puthucheary SD, Yasin R
    BMC Infect Dis, 2005 Feb 14;5:8.
    PMID: 15707504
    In Malaysia, Shigella spp. was reported to be the third commonest bacterial agent responsible for childhood diarrhoea. Currently, isolation of the bacterium and confirmation of the disease by microbiological and biochemical methods remain as the "gold standard". This study aimed to detect the prevalence of four Shigella virulence genes present concurrently, in randomly selected Malaysian strains via a rapid multiplex PCR (mPCR) assay.
    Matched MeSH terms: Antigens, Bacterial/genetics
  3. Distribution of Helicobacter pylori cagA, cagE and vacA in different ethnic groups in Kuala Lumpur, Malaysia
    Tan HJ, Rizal AM, Rosmadi MY, Goh KL
    J Gastroenterol Hepatol, 2005 Apr;20(4):589-94.
    PMID: 15836708
    There is a geographic variation in Helicobacter pylori (HP) genotypes and virulence factors. Cytotoxin associated genes A (cagA) and E (cagE), and certain vacuolating cytotoxin (vacA) genotypes are associated with peptic ulcer disease (PUD). There is also a different prevalence of PUD among different ethnic groups in Malaysia. The present study compared the distribution of vacA alleles and cagA and cagE status in three ethnic groups residing in Kuala Lumpur, Malaysia, and their association with clinical outcome.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  4. cagA gene variants in Malaysian Helicobacter pylori strains isolated from patients of different ethnic groups
    Ramelah M, Aminuddin A, Alfizah H, Isa MR, Jasmi AY, Tan HJ, et al.
    FEMS Immunol. Med. Microbiol., 2005 May 1;44(2):239-42.
    PMID: 15866222
    Helicobacter pylori infection of a distinct subtype of cagA may lead to different pathological manifestation. The aim of this study is to determine the presence of cagA gene and its variants in H. pylori infection among different ethnic groups and its effect on gastroduodenal diseases. Overall detection of cagA among the 205 clinical isolates of H. pylori was 94%. Variations in size of the 3' region of cagA gene were examined among 192 Malaysian H. pylori cagA-positive strains. Results showed that three cagA variants differing in fragment length of PCR products were detected and designated as type A (621-651bp), type B (732-735bp) and type C (525 bp). Although there was no association between any of the cagA subtypes with peptic ulcer disease (p>0.05), an association between cagA subtypes with a specific ethnic group was observed. Specific-cagA subtype A strains were predominantly isolated from Chinese compared to Malays and Indians (p<0.0005), and cagA subtype B strains were predominantly isolated from Malays and Indians compared to Chinese (p<0.05). The cagA type A strains of H. pylori is commonly found in the Chinese patients who have a higher risk of peptic ulcer disease, thus indicating that it could be used as an important clinical biomarker for a more severe infection.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  5. Intrabacterial proton-dependent CagA transport system in Helicobacter pylori
    Wu H, Nakano T, Daikoku E, Morita C, Kohno T, Lian HH, et al.
    J Med Microbiol, 2005 Dec;54(Pt 12):1117-1125.
    PMID: 16278423 DOI: 10.1099/jmm.0.46158-0
    Helicobacter pylori CagA modifies the signalling of host cells and causes gastric diseases. Although CagA is injected into gastric epithelial cells through the type IV secretion machinery, it remains unclear how CagA is transported towards the machinery in the bacterial cytoplasm. In this study, it was determined that the proton-dependent intracytoplasmic transport system correlates with the priming of CagA secretion from H. pylori. The cytotoxicity of neutral-pH- and acidic-pH-treated H. pylori was examined in the AGS cell line. The amount of phosphorylated CagA in AGS cells incubated with acidic-pH- and neutral-pH-treated H. pylori was determined by enzyme immunoassay and Western blot. The production of CagA and adherence of the treated bacteria were examined by enzyme immunoassay and light microscopy, respectively. To clarify how CagA is transported towards the inner membrane of the treated bacteria, the localization of CagA was analysed by immunoelectron microscopy. The proportion of hummingbird cells in the AGS cell line rapidly increased following the inoculation of acidic-pH-treated H. pylori but increased more slowly with neutral-pH-treated H. pylori, and the phenomenon correlated with the amount of phosphorylated CagA in AGS cells. CagA was densely localized near the inner membrane in the acidic-pH-treated bacterial cytoplasm, but this localization was not observed in the neutral-pH-treated bacterial cytoplasm, suggesting that CagA shifts from the centre to the peripheral portion of the cytoplasm as a result of an extracellular decrease in pH. This phenomenon depended on the presence of UreI, a proton-dependent urea channel, but not on the presence of urea. The pH treatments did not enhance CagA production or the adherence of the bacterium to AGS cells. The authors propose that H. pylori possesses a proton-dependent intracytoplasmic transport system that probably accelerates priming for CagA injection.
    Matched MeSH terms: Antigens, Bacterial/genetics
  6. Sequence analysis of the hypervariable regions of the 56 kDa immunodominant protein genes of Orientia tsutsugamushi strains in Malaysia
    Tay ST, Rohani YM, Ho TM, Shamala D
    Microbiol. Immunol., 2005;49(1):67-71.
    PMID: 15665455
    The DNA sequences encompassing two hypervariable regions, VD II and III of the 56 kDa immunodominant protein gene of 21 Malaysian strains of Orientia tsutsugamushi were determined. Two strains demonstrated a 100% DNA homology with the Gilliam prototype strain, and one with TH1817 strain and TA678 strain respectively. High percentages of DNA similarity (95-99%) were observed with Karp (4 strains), Gilliam (2 strains), TH1817 (4 strains), TC586 (3 strains) and TA763 (1 strain). The remaining strains demonstrated the highest DNA similarity with TA763 (1 strain, 89%), TA678 (1 strain, 86%) and TA686 (1 strain, 87%). Our study provides additional evidence on the existence and the genetic heterogeneity of TA strains of the Southeast Asia and their closely related strains in Malaysia.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  7. Role of Helicobacter pylori virulence factor and genotypes in non-ulcer dyspepsia
    Tan HJ, Rizal AM, Rosmadi MY, Goh KL
    J Gastroenterol Hepatol, 2006 Jan;21(1 Pt 1):110-5.
    PMID: 16706821
    The role of Helicobacter pylori (HP) in non-ulcer dyspepsia is debatable. Eradicating HP will help a small group of non-ulcer dyspeptic patients. However, it is unclear which subgroup of patients will benefit from eradication therapy. The aim of the present study was to compare the cagA and cagE status, as well as vacA genotypes, of HP in non-ulcer dyspeptic patients who responded successfully to eradication therapy compared with those patients who did not.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  8. Cloning, expression, and purification of recombinant protein from a single synthetic multivalent construct of Mycobacterium tuberculosis
    Fang CM, Zainuddin ZF, Musa M, Thong KL
    Protein Expr Purif, 2006 Jun;47(2):341-7.
    PMID: 16510294 DOI: 10.1016/j.pep.2005.12.007
    Tuberculosis remains a major infectious disease with over 8 million new cases and 2 million deaths annually. Therefore, a vaccine more potent than BCG is desperately needed. In this regard, an approximately 800 bp DNA encoding a mycobacterial synthetic gene designated as VacIII (containing ubiquitin gene UbGR and four immunogenic mycobacterial epitopes or genes of ESAT-6, Phos1, Hsp 16.3, and Mtb8.4) was sub-cloned into a bacterial expression vector of pRSET-B resulting in a 6 x His-VacIII fusion gene construction. This recombinant clone was over expressed in Escherichia coli BL-21 (DE-3). The expressed fusion protein was found almost entirely in the insoluble form (inclusion bodies) in cell lysate. The inclusion bodies were solubilized with 8M urea and the recombinant protein was purified by Ni-NTA column and dialyzed by urea gradient dialysis. This method produced a relatively high yield of recombinant VacIII protein and the cloned VacIII gene offers the potential development of other vaccine formats such as DNA vaccine and recombinant vaccine.
    Matched MeSH terms: Antigens, Bacterial/genetics
  9. Helicobacter pylori, ethnicity, and the gastroesophageal reflux disease spectrum: a study from the East
    Rajendra S, Ackroyd R, Robertson IK, Ho JJ, Karim N, Kutty KM
    Helicobacter, 2007 Apr;12(2):177-83.
    PMID: 17309756
    Ethnic differences in gastroesophageal reflux disease (GERD) and its complications as well as racial variations in the prevalence of Helicobacter pylori infection are well documented. Nevertheless, the association between reflux disease, H. pylori, and race has not been adequately explored.
    Matched MeSH terms: Antigens, Bacterial/genetics
  10. Fulltext Helicobacter pylori cagA gene variants in Malaysians of different ethnicity
    Mohamed R, Hanafiah A, Rose IM, Manaf MR, Abdullah SA, Sagap I, et al.
    Eur J Clin Microbiol Infect Dis, 2009 Jul;28(7):865-9.
    PMID: 19247698 DOI: 10.1007/s10096-009-0712-x
    We have defined DNA repeat variability in the 3'-terminus of the cagA gene of Helicobacter pylori strains from Malaysian patients of different ethnicities. We identified different alleles based on the EPIYA repeats. cagA types A-B-D and A-B-B-D are more similar to the sequence of Japanese strains, whereas cagA types A-B-C, A-B-C-C, A-B and A-C displayed similarity to strain 26695 sequences. A significant association was found between cagA genotypes and patients' ethnicity, with cagA type A-B-D being predominantly isolated from Chinese patients and cagA type A-B-C from Malays and Indians. Our data further corroborate the possibility that variant biological activity of CagA may affect the host specificity and/or pathogenicity of H. pylori.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  11. Distinct cagA EPIYA motifs are associated with ethnic diversity in Malaysia and Singapore
    Schmidt HM, Goh KL, Fock KM, Hilmi I, Dhamodaran S, Forman D, et al.
    Helicobacter, 2009 Aug;14(4):256-63.
    PMID: 19674129 DOI: 10.1111/j.1523-5378.2009.00684.x
    In vitro studies have shown that the biologic activity of CagA is influenced by the number and class of EPIYA motifs present in its variable region as these motifs correspond to the CagA phosphorylation sites. It has been hypothesized that strains possessing specific combinations of these motifs may be responsible for gastric cancer development. This study investigated the prevalence of cagA and the EPIYA motifs with regard to number, class, and patterns in strains from the three major ethnic groups within the Malaysian and Singaporean populations in relation to disease development.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  12. Clinical significance of Helicobacter pylori cagA and iceA genotype status
    Amjad N, Osman HA, Razak NA, Kassian J, Din J, bin Abdullah N
    World J Gastroenterol, 2010 Sep 21;16(35):4443-7.
    PMID: 20845512
    AIM: To study the presence of Helicobacter pylori (H. pylori) virulence factors and clinical outcome in H. pylori infected patients.

    METHODS: A prospective analysis of ninety nine H. pylori-positive patients who underwent endoscopy in our Endoscopy suite were included in this study. DNA was isolated from antral biopsy samples and the presence of cagA, iceA, and iceA2 genotypes were determined by polymerase chain reaction and a reverse hybridization technique. Screening for H. pylori infection was performed in all patients using the rapid urease test (CLO-Test).

    RESULTS: From a total of 326 patients who underwent endoscopy for upper gastrointestinal symptoms, 99 patients were determined to be H. pylori-positive. Peptic ulceration was seen in 33 patients (33%). The main virulence strain observed in this cohort was the cagA gene isolated in 43 patients. cagA was associated with peptic ulcer pathology in 39.5% (17/43) and in 28% (16/56) of non-ulcer patients. IceA1 was present in 29 patients (29%) and iceA2 in 15 patients (15%). Ulcer pathology was seen in 39% (11/29) of patients with iceA1, while 31% (22/70) had normal findings. The corresponding values for iceA2 were 33% (5/15) and 33% (28/84), respectively.

    CONCLUSION: Virulence factors were not common in our cohort. The incidence of factors cagA, iceA1 and iceA2 were very low although variations were noted in different ethnic groups.

    Matched MeSH terms: Antigens, Bacterial/genetics*
  13. DNA vaccine encoding avian influenza virus H5 and Esat-6 of Mycobacterium tuberculosis improved antibody responses against AIV in chickens
    Oveissi S, Omar AR, Yusoff K, Jahanshiri F, Hassan SS
    Comp Immunol Microbiol Infect Dis, 2010 Dec;33(6):491-503.
    PMID: 19781778 DOI: 10.1016/j.cimid.2009.08.004
    The H5 gene of avian influenza virus (AIV) strain A/chicken/Malaysia/5744/2004(H5N1) was cloned into pcDNA3.1 vector, and Esat-6 gene of Mycobacterium tuberculosis was fused into downstream of the H5 gene as a genetic adjuvant for DNA vaccine candidates. The antibody level against AIV was measured using enzyme-linked immunosorbent assay (ELISA) and haemagglutination inhibition (HI) test. Sera obtained from specific-pathogen-free chickens immunized with pcDNA3.1/H5 and pcDNA3.1/H5/Esat-6 demonstrated antibody responses as early as 2 weeks after the first immunization. Furthermore, the overall HI antibody titer in chickens immunized with pcDNA3.1/H5/Esat-6 was higher compared to the chickens immunized with pcDNA3.1/H5 (p<0.05). The results suggested that Esat-6 gene of M. tuberculosis is a potential genetic adjuvant for the development of effective H5 DNA vaccine in chickens.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  14. Structural and functional studies of a 50 kDa antigenic protein from Salmonella enterica serovar Typhi
    Choong YS, Lim TS, Chew AL, Aziah I, Ismail A
    J Mol Graph Model, 2011 Apr;29(6):834-42.
    PMID: 21371926 DOI: 10.1016/j.jmgm.2011.01.008
    The high typhoid incidence rate in developing and under-developed countries emphasizes the need for a rapid, affordable and accessible diagnostic test for effective therapy and disease management. TYPHIDOT®, a rapid dot enzyme immunoassay test for typhoid, was developed from the discovery of a ∼50 kDa protein specific for Salmonella enterica serovar Typhi. However, the structure of this antigen remains unknown till today. Studies on the structure of this antigen are important to elucidate its function, which will in turn increase the efficiency of the development and improvement of the typhoid detection test. This paper described the predictive structure and function of the antigenically specific protein. The homology modeling approach was employed to construct the three-dimensional structure of the antigen. The built structure possesses the features of TolC-like outer membrane protein. Molecular docking simulation was also performed to further probe the functionality of the antigen. Docking results showed that hexamminecobalt, Co(NH(3))(6)(3+), as an inhibitor of TolC protein, formed favorable hydrogen bonds with D368 and D371 of the antigen. The single point (D368A, D371A) and double point (D368A and D371A) mutations of the antigen showed a decrease (single point mutation) and loss (double point mutations) of binding affinity towards hexamminecobalt. The architecture features of the built model and the docking simulation reinforced and supported that this antigen is indeed the variant of outer membrane protein, TolC. As channel proteins are important for the virulence and survival of bacteria, therefore this ∼50 kDa channel protein is a good specific target for typhoid detection test.
    Matched MeSH terms: Antigens, Bacterial/genetics
  15. Fulltext Travel-related Streptococcal toxic shock syndrome caused by emm type 78 Streptococcus pyogenes
    Tappe D, Schulze MH, van der Linden M, Ziegler U, Müller A, Stich A
    J Clin Microbiol, 2011 Aug;49(8):3094-5.
    PMID: 21632896 DOI: 10.1128/JCM.02623-10
    Streptococcal toxic shock syndrome is a serious health problem in developed and developing countries. We here report a case of severe protracted disease after a minor skin infection in a young traveler returning from West Malaysia which was caused by an unusual emm-type strain harboring speG and smeZ superantigen genes.
    Matched MeSH terms: Antigens, Bacterial/genetics
  16. Fulltext Variant of Helicobacter pylori CagA proteins induce different magnitude of morphological changes in gastric epithelial cells
    Alfizah H, Ramelah M
    Malays J Pathol, 2012 Jun;34(1):29-34.
    PMID: 22870595 MyJurnal
    Infection with Helicobacter pylori cagA-positive strains is associated with gastroduodenal diseases. The CagA protein is injected into gastric epithelial cells and supposedly induces morphological changes termed the 'hummingbird phenotype', which is associated with scattering and increased cell motility. The molecular mechanisms leading to the CagA-dependent morphological changes are only partially known. The present study was carried out to investigate the effect of CagA variants on the magnitude of gastric epithelial cell morphological changes. Recombinant 3' terminal domains of cagA were cloned and expressed in a gastric epithelial cell line and the hummingbird phenotype was quantified by microscopy. The 3' region of the cagA gene of Malaysian H. pylori isolates showed six sub-genotypes that differed in the structural organization of the EPIYA repeat sequences. The percentage of hummingbird cells induced by CagA increased with duration of transfection. The hummingbird phenotype was observed to be more pronounced when CagA with 4 EPIYA motifs rather than 3 or 2 EPIYA motifs was produced. The activity of different CagA variants in the induction of the hummingbird phenotype in gastric epithelial cells depends at least in part on EPIYA motif variability. The difference in CagA genotypes might influence the potential of individual CagAs to cause morphological changes in host cells. Depending on the relative exposure of cells to CagA genotypes, this may contribute to the various disease outcomes caused by H. pylori infection in different individuals.
    Matched MeSH terms: Antigens, Bacterial/genetics*
  17. Association of Malaysian Helicobacter pylori virulence polymorphisms with severity of gastritis and patients' ethnicity
    Alfizah H, Ramelah M, Rizal AM, Anwar AS, Isa MR
    Helicobacter, 2012 Oct;17(5):340-9.
    PMID: 22967117 DOI: 10.1111/j.1523-5378.2012.00956.x
    Polymorphisms of Helicobacter pylori cagA and vacA genes do exist and may contribute to differences in H. pylori infection and gastroduodenal diseases among races in the Malaysian population. This study was conducted to characterize the polymorphisms in H. pylori cagA and vacA in Malaysian population.
    Matched MeSH terms: Antigens, Bacterial/genetics
  18. Fulltext A real-time loop-mediated isothermal amplification assay for rapid detection of Shigella species
    Liew PS, Teh CS, Lau YL, Thong KL
    Trop Biomed, 2014 Dec;31(4):709-20.
    PMID: 25776596 MyJurnal
    Shigellosis is a foodborne illness caused by the genus Shigella and is an important global health issue. The development of effective techniques for rapid detection of this pathogen is essential for breaking the chain of transmission. Therefore, we have developed a novel loop-mediated isothermal amplification (LAMP) assay targeting the invasion plasmid antigen H (ipaH) gene to rapidly detect Shigella species. This assay could be performed in 90 min at an optimal temperature of 64ºC, with endpoint results visualized directly. Notably, the method was found to be more sensitive than conventional PCR. Indeed, the detection limit for the LAMP assay on pure bacterial cultures was 5.9 x 10(5) CFU/ml, while PCR displayed a limit of 5.9 x 10(7) CFU/ml. In spiked lettuce samples, the sensitivity of the LAMP assay was 3.6 x 10(4) CFU/g, whereas PCR was 3.6 x 10(5) CFU/g. Overall, the assay accurately identified 32 Shigella spp. with one enteroinvasive Escherichia coli displaying positive reaction while the remaining 32 non-Shigella strains tested were negative.
    Matched MeSH terms: Antigens, Bacterial/genetics
  19. Fulltext Multi-isotype antibody responses against the multimeric Salmonella Typhi recombinant hemolysin E antigen
    Ong EB, Ignatius J, Anthony AA, Aziah I, Ismail A, Lim TS
    Microbiol. Immunol., 2015 Jan;59(1):43-7.
    PMID: 25399538 DOI: 10.1111/1348-0421.12211
    The detection and measurement of different antibody isotypes in the serum provide valuable indicators of the different stages of typhoid infection. Here, the ability of S. Typhi recombinant hemolysin E (HlyE) to detect multi-isotype antibody responses in sera of patients with typhoid and paratyphoid A was investigated using an indirect antibody immunoassay. Nanogram amounts of HlyE were found to be sufficient for detection of IgG and IgA isotypes and, in a study of individuals' sera (n = 100), the immunoassay was able to distinguish between typhoid and non-typhoid sera. The overall sensitivity, specificity and efficiency of the ELISA were 70% (39/56), 100% (44/44) and 83% respectively.
    Matched MeSH terms: Antigens, Bacterial/genetics
  20. Fulltext Crystal structure of an antigenic outer-membrane protein from Salmonella Typhi suggests a potential antigenic loop and an efflux mechanism
    Guan HH, Yoshimura M, Chuankhayan P, Lin CC, Chen NC, Yang MC, et al.
    Sci Rep, 2015 Nov 13;5:16441.
    PMID: 26563565 DOI: 10.1038/srep16441
    ST50, an outer-membrane component of the multi-drug efflux system from Salmonella enterica serovar Typhi, is an obligatory diagnostic antigen for typhoid fever. ST50 is an excellent and unique diagnostic antigen with 95% specificity and 90% sensitivity and is used in the commercial diagnosis test kit (TYPHIDOT(TM)). The crystal structure of ST50 at a resolution of 2.98 Å reveals a trimer that forms an α-helical tunnel and a β-barrel transmembrane channel traversing the periplasmic space and outer membrane. Structural investigations suggest significant conformational variations in the extracellular loop regions, especially extracellular loop 2. This is the location of the most plausible antibody-binding domain that could be used to target the design of new antigenic epitopes for the development of better diagnostics or drugs for the treatment of typhoid fever. A molecule of the detergent n-octyl-β-D-glucoside is observed in the D-cage, which comprises three sets of Asp361 and Asp371 residues at the periplasmic entrance. These structural insights suggest a possible substrate transport mechanism in which the substrate first binds at the periplasmic entrance of ST50 and subsequently, via iris-like structural movements to open the periplasmic end, penetrates the periplasmic domain for efflux pumping of molecules, including poisonous metabolites or xenobiotics, for excretion outside the pathogen.
    Matched MeSH terms: Antigens, Bacterial/genetics
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