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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*; Antigens, Bacterial/metabolism; Antigens, Bacterial/chemistry*
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; Antigens, Bacterial/metabolism; Antigens, Bacterial/chemistry*
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/biosynthesis; Antigens, Bacterial/genetics; Antigens, Bacterial/chemistry
Fulltext Helicobacter pylori Virulence Factor Cytotoxin-Associated Gene A (CagA)-Mediated Gastric Pathogenicity

Ansari S, Yamaoka Y

Int J Mol Sci, 2020 Oct 08;21(19).
PMID: 33050101 DOI: 10.3390/ijms21197430

Helicobacter pylori causes persistent infection in the gastric epithelium of more than half of the world's population, leading to the development of severe complications such as peptic ulcer diseases, gastric cancer, and gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Several virulence factors, including cytotoxin-associated gene A (CagA), which is translocated into the gastric epithelium via the type 4 secretory system (T4SS), have been indicated to play a vital role in disease development. Although infection with strains harboring the East Asian type of CagA possessing the EPIYA-A, -B, and -D sequences has been found to potentiate cell proliferation and disease pathogenicity, the exact mechanism of CagA involvement in disease severity still remains to be elucidated. Therefore, we discuss the possible role of CagA in gastric pathogenicity.

Matched MeSH terms: Antigens, Bacterial/genetics*; Antigens, Bacterial/metabolism; Antigens, Bacterial/chemistry
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; Antigens, Bacterial/metabolism; Antigens, Bacterial/chemistry*
A 35 kDa antigenic protein from Shigella flexneri: in silico structural and functional studies

Yung-Hung RL, Ismail A, Lim TS, Choong YS

Biochem Biophys Res Commun, 2011 Nov 18;415(2):229-34.
PMID: 21982766 DOI: 10.1016/j.bbrc.2011.09.116

Shigella flexneri serotype 2a is a major public health concern in the developing and under-developed countries which contributes to shigellosis endemic and mortality. Thus, there is an urgent need for a rapid diagnostic test for effective therapy and disease management. Previous study showed that a ∼35 kDa antigenic protein from S. flexneri is a potential biomarker. We therefore modelled the three-dimensional structure of the antigen to probe its functionality which could aid in the development of an antigen-based diagnostic. Results showed that the antigen is a transmembrane protein consists of OmpA and OmpA-like domains. The OmpA domain is a beta-barrel embedded in the outer membrane with four surface-exposed extracellular loops. The OmpA-like domain is linked to the OmpA domain with a 17 amino acids linker and located in the periplasmic. Docking of peptidoglycan into the groove of OmpA-like domain might help in catalyzing the bacterial cell wall formation. Both domains are expected to be involved in the virulence, structural stability, pathogenesis and survival of Shigella thus made the 35 kDa protein a suitable shigellosis diagnostic biomarker. This structural elucidation will also enable a better identification of the epitope regions for the development of specific binders to the 35 kDa antigen.

Matched MeSH terms: Antigens, Bacterial/chemistry*
Burkholderia thailandensis whole cell antigen cross-reacts with B. pseudomallei antibodies from patients with melioidosis in an immunofluorescent assay

Puthucheary SD, Anuar AS, Tee TS

Southeast Asian J. Trop. Med. Public Health, 2010 Mar;41(2):395-400.
PMID: 20578523

An immunofluorescent assay (IFAT) using whole cell antigen derived from Burkholderia thailandensis used for detection of total antibodies to Burkholderia pseudomallei, was found to compare favorably with a previous published report on a B. pseudomallei IFAT assay. At a 1:20 cut-off titer, the assay had high sensitivity (98.9%) and satisfactory specificity (92.3%), when tested against sera from 94 patients suspected of melioidosis. Sera from 12 patients with culture proven melioidosis gave absolute concordance with the 2 test antigens. No sera from 50 blood donors had a titer of > or =20. Cross-reactivity with patients' sera positive for Chlamydia, Mycoplasma, Legionella and typhoid was not observed, except for 3 sera from typhus patients and one from a patient with leptospirosis. The major advantage of this assay is that the cultivation and preparation of B. thailandensis as antigen can be carried out in any laboratory with basic microbiological set-up. The serodiagnosis of melioidosis can be made safe for medical laboratory personnel, particularly in B. pseudomallei endemic regions.

Matched MeSH terms: Antigens, Bacterial/immunology*
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*
Prevalence of antibodies to Pseudomonas pseudomallei exotoxin and whole cell antigens in military personnel in Sabah and Sarawak, Malaysia

Embi N, Suhaimi A, Mohamed R, Ismail G

Microbiol. Immunol., 1992;36(8):899-904.
PMID: 1474938

Sera from 420 military personnel serving in Sabah and Sarawk, Malaysia, were tested for antibodies to Pseudomonas pseudomallei exotoxin and whole cell antigens by enzyme-linked immunosorbent assay procedure (ELISA). Data showed that 54.4% of serum samples were positive for antibodies to P. pseudomallei exotoxin and 65.7% were positive for antibodies to the whole cell antigens. Samples gave much lower titers for anti-exotoxin antibodies compared to titers against crude whole cell antigens. The incidence of antibody to exotoxin was highest in the age groups ranging from 26 to 32 years, where the positive rates were higher than 40% and 30% for military personnel serving in Sarawak and Sabah, respectively.

Matched MeSH terms: Antigens, Bacterial/blood*
Limitations of serodiagnosis in chlamydial genital tract infections

Ngeow YF

Ann Acad Med Singap, 1996 Mar;25(2):300-4.
PMID: 8799029

Infection with Chlamydia trachomatis results in the formation of a variety of antibodies with group, species, subspecies and serovarspecificity. Sera from patients with genital tract infections often show broad reactivity in serological tests. This may be due to the presence of cross-reacting antibodies, repeated infections by different serotypes or concurrent genital and respiratory infections by different chlamydial species. Other factors contributing to difficulties in interpretation include how antibody titres in acute mucosal infections, the occurrence of latent infections and reactivations, and the persistence of IgG which does not allow the differentiation of past from current infections. For these reasons, serology alone is inadequate for the diagnosis of uncomplicated lower genital tract infections. In upper genital tract infections, however, because of difficulties with sampling from the infected site, a positive serology may be the only indications of chlamydial involvement. This paper discusses the principles of chlamydial antibody assays, difficulties with their interpretation and their role in the diagnosis of upper and lower genital tract infections.

Matched MeSH terms: Antigens, Bacterial/analysis*
Preliminary evaluation of the INDX DIP-S-TICKS with positive rickettsial samples in Malaysia

Koay AS, Cheong YM

Malays J Pathol, 1993 Jun;15(1):69-73.
PMID: 8277794

Forty-four serum samples of various reactivities to rickettsial antigens demonstrated by the indirect immunoperoxidase technique were tested with INDX Dip-S-Ticks (INDX Integrated Diagnostics Inc., USA) Kit for the detection of tick borne diseases. The kit utilised Rickettsia rickettsii the causative agent of Rocky Mountain spotted fever (RMSF) as antigens. The samples positive for endemic typhus were also tested against R. typhi, the agent for endemic typhus by the same method. The aim of this study was to determine the extent of cross-reactivity of R. rickettsii with rickettsial infections in Malaysia. Nine out of 12 tick typhus, 4 out of 10 scrub typhus and 4 out of 12 endemic typhus samples cross reacted with R. rickettsii. Ten out of 12 endemic samples were positive with R. typhi by the same method. From the study, we concluded that the INDX Dip-S-Ticks Kit can be used as a rapid screening test to detect endemic and tick-borne rickettsial infections in Malaysia but a second serological test is strongly recommended on all weakly reactive cases.

Matched MeSH terms: Antigens, Bacterial/analysis*
Rapid serodiagnosis of typhoid fever by dot enzyme immunoassay in an endemic area

Choo KE, Oppenheimer SJ, Ismail AB, Ong KH

Clin Infect Dis, 1994 Jul;19(1):172-6.
PMID: 7948526

A dot enzyme immunoassay (EIA) using 50-kD outer-membrane proteins (OMPs) of Salmonella typhi was compared with the Widal test for the serodiagnosis of typhoid fever in 109 febrile children admitted to a hospital in an endemic area. In the culture-positive typhoid group, the initial dot EIA was positive in 40 of 42 cases and the initial Widal test was positive in 41. In the culture-negative clinical typhoid group, both the dot EIA and the Widal test were positive in 17 of 18 cases. In the nontyphoidal fever group, the dot EIA was negative in all of 49 cases and the Widal test was negative in 44. With culture used as the gold standard, the dot EIA is as sensitive as the Widal test (95% vs. 98%), has a similar high negative predictive value (96% vs. 98%), and is more specific (75% vs. 67%). In addition, the dot EIA offers the advantages of simplicity, speed, early diagnosis, economy, and flexibility (i.e., other diagnostic tests can be conducted simultaneously).

Matched MeSH terms: Antigens, Bacterial/immunology*
Fulltext A preliminary study on the use of counterimmunoelectrophoresis and coagglutination methods in the diagnosis of bacterial meningitis

Cheong YM, Jegathesan M, Lo SB

Med J Malaysia, 1984 Mar;39(1):38-41.
PMID: 6513838

The usefulness of counterimmunoelectrophoresis (CIEP) and coagglutination (COAG) methods in the diagnosis of bacterial meningitis was evaluated. Out of the 31 cerebrospinal fluid (CSF) specimens which had a cell count of >5 x10^6 wbc/l and were negative on gram stain and culture, pneumococcal antigens were detected in four specimens and Haemophilus influenzae type b antigen was detected in one specimen by both the methods. No false positives were detected in 10 specimens obtained from cases of febrile fits whose CSF showed no evidence of meningitis. One CSF sample, from which Klebsiella spp. was isolated, cross reacted with the meningococcal polyvalent group A-D antiserum in the CIEP test. From this study we found that these methods are rapid, simple and useful adjunctive tests In the diagnosis of bacterial meningitis, especially in the partially treated cases.

Matched MeSH terms: Antigens, Bacterial/analysis*
Demonstration of an antigenic protein specific for Salmonella typhi

Ismail A, Hai OK, Kader ZA

Biochem Biophys Res Commun, 1991 Nov 27;181(1):301-5.
PMID: 1958200

Current studies were undertaken to determine the presence of a specific antigenic protein on the outer membrane of Salmonella typhi. Immunoblot analysis using sera from patients with fevers revealed that the 50 kD band was specifically recognized only by typhoid sera. The 50 kD band located on the outer membrane is protein by nature and is not a Vi (capsular), dH (flagellar), or O9 (somatic) antigen of S. typhi. These results indicate the usefulness of the specific antigen in the development of a serodiagnostic test for typhoid fever since antibodies of both the IgM and IgG class responses were obtained.

Matched MeSH terms: Antigens, Bacterial/analysis*
Mycobacterium leprae reactive T cell clones from lepromatous leprosy patients after prolonged dapsone chemotherapy

Gill HK, Ridley DS, Ganesan J, Mustafa AS, Rees RJ, Godal T

Lepr Rev, 1990 Mar;61(1):25-31.
PMID: 2181222

The proliferative responses of peripheral blood mononuclear cells (PBMC) to Mycobacterium leprae and BCG were studied in two groups of leprosy patients: a group of 8 lepromatous patients who had been on treatment for more than 20 years (TLL) and a group of 8 untreated lepromatous leprosy patients (ULL). The mean response to M. leprae of the TLL group was 6195 cpm with 5 of the 8 patients responding positively. The mean response to M. leprae of the ULL group was 617 cpm, with only 1 patient showing a positive response. The corresponding proliferative responses to BCG were 19,908 cpm in the TLL group and 7908 in the ULL group. Thirteen M. leprae reactive clones were established from 2 TLL patients and 5 M. leprae reactive clones were established from 2 tuberculoid leprosy patients. Seven of these clones, 4 from the TLL patients and 3 from the tuberculoid (TT) patients could be studied further. Three of the TLL clones responded specifically to M. leprae, while one of the clones exhibited a broad cross-reactivity to other mycobacteria. All of these clones were of the CD4+CD8- phenotype. Our findings suggest that responsiveness to M. leprae can be detected in vitro in a proportion of LL patients who have undergone prolonged chemotherapy, and that this response involves M. leprae reactive CD8+CD8- T cells, of which some appear to be specific to M. leprae.

Matched MeSH terms: Antigens, Bacterial/immunology
Antigenic analysis by direct immunofluorescence of 114 isolates of Rickettsia tsutsugamushi recovered from febrile patients in rural Malaysia

Shirai A, Robinson DM, Brown GW, Gan E, Huxsoll DL

Jpn. J. Med. Sci. Biol., 1979 Dec;32(6):337-44.
PMID: 120901

One hundred and fourteen Rickettsia tsutsugamushi isolates, recovered from febrile patients in central Peninsular Malaysia, were antigenically analyzed by direct immunofluorescence using eight prototype strains. Twenty-nine antigenic types were detected. The TA763, TA716, Karp and TA686 strains were the most common and occurred singly or in combination with each other or other strains in 86% of the isolates.

Matched MeSH terms: Antigens, Bacterial/analysis*
Fulltext In Silico identification of M. TB proteins with diagnostic potential

Calero R, Mirabal M, Bouza J, Guzmán MV, Carrillo H, López Y, et al.

BMC Immunol, 2013;14 Suppl 1:S9.
PMID: 23458073 DOI: 10.1186/1471-2172-14-S1-S9

TB, caused by Mycobacterium tuberculosis (MTB), is one of the major global infectious diseases. For the pandemic control, early diagnosis with sensitive and specific methods is fundamental. With the advent of bioinformatics' tools, the identification of several proteins involved in the pathogenesis of TB (TB) has been possible. In the present work, the MTB genome was explored to look for molecules with possible antigenic properties for their evaluation as part of new generation diagnostic kits based on the release of cytokines. Seven proteins from the MTB proteome and some of their combinations suited the computational test and the results suggested their potential use for the diagnosis of infection in the following population groups: Cuba, Mexico, Malaysia and sub-Saharan Africa. Our predictions were performed using public bioinformatics tools plus three computer programs, developed by our group, to facilitate information retrieval and processing.

Matched MeSH terms: Antigens, Bacterial/immunology
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*
Mimotopes of the Vi antigen of Salmonella enterica serovar typhi identified from phage display peptide library

Tang SS, Tan WS, Devi S, Wang LF, Pang T, Thong KL

Clin Diagn Lab Immunol, 2003 Nov;10(6):1078-84.
PMID: 14607870

The capsular polysaccharide Vi antigen (ViCPS) is an essential virulence factor and also a protective antigen of Salmonella enterica serovar Typhi. A random 12-mer phage-displayed peptide library was used to identify mimotopes (epitope analogues) of this antigen by panning against a ViCPS-specific monoclonal antibody (MAb) ATVi. Approximately 75% of the phage clones selected in the fourth round carried the peptide sequence TSHHDSHGLHRV, and the rest of the clones harbored ENHSPVNIAHKL and other related sequences. These two sequences were also obtained in a similar panning process by using pooled sera from patients with a confirmed diagnosis of typhoid fever, suggesting they mimic immunodominant epitopes of ViCPS antigens. Binding of MAb ATVi to the mimotopes was specifically blocked by ViCPS, indicating that they interact with the same binding site (paratope) of the MAb. Data and reagents generated in this study have important implications for the development of peptide-base diagnostic tests and peptide vaccines and may also provide a better understanding of the pathogenesis of typhoid fever.

Matched MeSH terms: Antigens, Bacterial/immunology*
Development of an immunochromatographic lateral flow dipstick for the detection of Mycobacterium tuberculosis 16 kDa antigen (Mtb-strip)

Mohd Amiruddin MN, Ang GY, Yu CY, Falero-Diaz G, Otero O, Reyes F, et al.

J Microbiol Methods, 2020 09;176:106003.
PMID: 32702386 DOI: 10.1016/j.mimet.2020.106003

Mycobacterium tuberculosis (Mtb) is a pathogenic bacterium that causes tuberculosis (TB). This contagious disease remains a severe health problem in the world. The disease is transmitted via inhalation of airborne droplets carrying Mtb from TB patients. Early detection of the disease is vital to prevent transmission of the infection to people in close contact with the patients. To date, there is a need of a simple, rapid, sensitive and specific diagnostic test for TB. Previous studies showed the potential of Mtb 16 kDa antigen (Ag16) in TB diagnosis. In this study, lateral flow immunoassay, also called simple strip immunoassay or immunochromatographic test (ICT) for detection of Ag16 was developed (Mtb-strip) and assessed as a potential rapid TB diagnosis method. A monoclonal antibody against Ag16 was optimized as the capturing and detection antibody on the Mtb-strip. Parameters affecting the performance of the Mtb-strip were also optimized before a complete prototype was developed. Analytical sensitivity showed that Mtb-strip was capable to detect as low as 125 ng of purified Ag16. The analytical sensitivity of Mtb-strip suggests its potential usefulness in different clinical applications.

Matched MeSH terms: Antigens, Bacterial/immunology*

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