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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: Antibodies, Bacterial/immunology*
Fulltext Evaluation of recombinant antigens for diagnosis of melioidosis

Allwood EM, Logue CA, Hafner GJ, Ketheesan N, Norton RE, Peak IR, et al.

FEMS Immunol. Med. Microbiol., 2008 Oct;54(1):144-53.
PMID: 18657105 DOI: 10.1111/j.1574-695X.2008.00464.x

Burkholderia pseudomallei, the causative agent of melioidosis, is endemic to Southeast Asia and northern Australia. Clinical manifestations of the disease are diverse, ranging from chronic localized infection to acute septicaemia, with death occurring within 24-48 h after the onset of symptoms. Definitive diagnosis of melioidosis involves bacterial culture and identification, with results obtained within 3-4 days. This delayed diagnosis is a major contributing factor to high mortality rates. Rapid diagnosis is vital for successful management of the disease. This study describes the purification and evaluation of three recombinant antigenic proteins, BPSL0972, BipD and OmpA from B. pseudomallei 08, for their potential in the serodiagnosis of melioidosis using an indirect enzyme-linked immunosorbent assay (ELISA) method. The recombinant proteins were evaluated using 74 serum samples from culture-confirmed melioidosis patients from Malaysia, Thailand and Australia. In addition, 62 nonmelioidosis controls consisting of serum samples from clinically suspected melioidosis patients (n=20) and from healthy blood donors from an endemic region (n=18) and a nonendemic region (n=24) were included. The indirect ELISAs using BipD and BPSL0972 as antigens demonstrated poor to moderate sensitivities (42% and 51%, respectively) but good specificity (both 100%). In contrast, the indirect ELISA using OmpA as an antigen achieved 95% sensitivity and 98% specificity. These results highlight the potential for OmpA to be used in the serodiagnosis of melioidosis in an endemic area.

Matched MeSH terms: Antibodies, Bacterial/blood
Thoughts about populations with unexpected low prevalences of Helicobacter pylori infection

Graham DY, Yamaoka Y, Malaty HM

Trans R Soc Trop Med Hyg, 2007 Sep;101(9):849-51.
PMID: 17658569

Helicobacter pylori is one of the few remaining major pathogens that accompanied humans on their travels from Africa. A recently published study reports the unexpected finding of a low H. pylori prevalence among pregnant women in Zanzibar (Farag, T.H., Stolzfus, R.J., Khalfan, S.S., Tielsch, J.M., 2007. Unexpectedly low prevalence of Helicobacter pylori infection among pregnant women on Pemba Island, Zanzibar. Trans. R. Soc. Trop. Med. Hyg. 101). The apparent epidemiology of higher prevalence with higher socioeconomic status and decrease with age are unprecedented. As with many 'unexpected' events, a search of the literature reveals evidence of low prevalence populations in Java and Malaysia, with clues dating back to the mid-twentieth century. Why some populations apparently lost H. pylori infection remains an open question. However, the tools needed to resolve the dilemma are readily available and we hope investigators will soon rise to the challenge.

Matched MeSH terms: Antibodies, Bacterial/analysis*
The induction of oral tolerance to Actinomyces viscosus in mice

Sosroseno W, Bird PS, Gemmell E, Seymour GJ

Oral Dis, 2006 Jul;12(4):387-94.
PMID: 16792724

To determine whether oral tolerance with the oral bacterium Actinomyces viscosus was inducible in mice.

Matched MeSH terms: Antibodies, Bacterial/blood
The use of the indirect immunoperoxidase test for the serodiagnosis of rickettsial diseases in Malaysia

Tay ST, Rohani MY

Southeast Asian J. Trop. Med. Public Health, 2002 Jun;33(2):314-20.
PMID: 12236431

The indirect immunoperoxidase (HP) test has been used extensively in most government hospitals in Malaysia for the serodiagnosis of scrub typhus, murine typhus and tick typhus during the 1990s. The test was used to determine the IgG and IgM antibody titers in patients' sera for three rickettsial species, ie Orientia tsutsugamushi OT; the causative agent of scrub typhus), Rickettsia typhi (RT; the causative agent of murine typhus), and TT118 spotted fever group rickettsiae (TT; the causative agent of tick typhus). The serological findings obtained from Malaysian hospitals using the IIP test (1994-1999) were analyzed. During the six-year period, a total of 61,501 patients' sera were tested, of which 9.6%, 10.5%, and 12.9% had antibody (IgG and/or IgM of > or = 1:50) for OT, RT and TT respectively. A total of 8.6%, 9.8%, and 9.7% of sera had IgG antibody of > or = 1:50 for OT, RT, and TT respectively, indicating past infection. A total of 3.4%, 3.8%, and 6.4 % of sera had IgM antibody of > or = 1:50 for OT, RT, and TT respectively, indicating recent infection. A total of 2,986 (4.9%), 1,882 (3.1%), and 1,574 (2.6%) of sera had IgG and/or IgM antibody titers of > or = 1:400 for OT, RT, and TT respectively, suggesting active rickettsial infection. The seropositivity rates of OT, RT and TT varied according to geographical locations. While the seropositivity of OT remained constant during the six-year period, a reduction in the seropositivity of both RT and TT was noted during recent years. The serological findings reflect the endemicity of rickettsial diseases, including tick typhus, and endemic typhus in various parts of Malaysia. Awareness of these diseases by health and medical staff and by the general public is important if the mortality and morbidity associated with scrub typhus, tick typhus, and murine typhus in Malaysia, are to be reduced.

Matched MeSH terms: Antibodies, Bacterial/blood
Fulltext Comparison of safety and immunogenicity of a Vi polysaccharide typhoid vaccine with a whole-cell killed vaccine in Malaysian Air Force recruits

Panchanathan V, Kumar S, Yeap W, Devi S, Ismail R, Sarijan S, et al.

Bull World Health Organ, 2001;79(9):811-7.
PMID: 11584728

To carry out a comparative study of the safety and immunogenicity of Vi polysaccharide vaccine against whole-cell killed (WCK) typhoid vaccine.

Matched MeSH terms: Antibodies, Bacterial/blood
Fulltext Serological evidence of natural infection of wild rodents (Rattus spp and Tupaia glis) with rickettsiae in Malaysia

Tay ST, Kaewanee S, Ho TM, Rohani MY, Devi S

Southeast Asian J. Trop. Med. Public Health, 1998 Sep;29(3):560-2.
PMID: 10437957
Matched MeSH terms: Antibodies, Bacterial/isolation & purification*
Characterization of immune response and duration of protection in buffaloes immunized with haemorrhagic septicaemia vaccines

Chandrasekaran S, Kennett L, Yeap PC, Muniandy N, Rani B, Mukkur TK

Vet Microbiol, 1994 Aug 01;41(3):213-9.
PMID: 7975147

Two of the three buffaloes immunized with a non-adjuvanted broth bacterin were found to be protected against experimental challenge at 6 weeks but not at 3 months post-challenge. Similarly all buffaloes (4/4) immunized with alum-precipitated vaccine were protected at 6 months but only 1 of the 2 vaccinated animals were protected at 12 months post-immunization. On the other hand, buffaloes immunized with an oil adjuvant and a double emulsion vaccine were completely protected at 12 months post-immunization. Statistically significant differences between immunized versus non-immune animals became evident at 3 months post-immunization, although analysis of cumulative antibody titres of pre-challenge sera of vaccinated buffaloes surviving versus those succumbing to experimental challenge revealed significant by higher antibody titres in the former as compared to the latter group. These results suggested that there was a relationship between ELISA antibody titres and active protection in buffaloes. There also appeared to be a relationship between cutaneous delayed-type hypersensitivity and active protection in buffaloes. Preliminary analysis of the antibody isotype distribution in the pre-challenge sera of 2 buffaloes vaccinated with the oil adjuvant vaccine revealed predominance of IgG1 and IgG2 subclasses whose role in protection against haemorrhagic septicaemia was not eludicated.

Matched MeSH terms: Antibodies, Bacterial/blood
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: Antibodies, Bacterial/blood*
Fulltext Development and preliminary evaluation of an IgM dot-immunobinding assay (IgM-DIA) for rapid serodiagnosis of scrub typhus infection

Koay AS, Tay ST, Cheong YM, Yasin RM

Southeast Asian J. Trop. Med. Public Health, 1995 Jun;26(2):350-3.
PMID: 8629074

An IgM dot-immunobinding assay (IgM-DIA) was developed for the diagnosis of scrub typhus infection. The whole cell antigens of Karp, Kato and Gilliam strains of Rickettsia tsutsugamushi were immobilized onto nitrocellulose paper and reacted with patients sera. The presence of IgM R. tsutsugamushi specific antibody in the patient sera could be detected by the observation of a visible brown dot on the nitrocellulose paper. The IgM-DIA has a sensitivity of 90.4% and specificity of 81.4% as compared to the indirect immunoperoxidase test. The IgM-DIA is rapid, simple, cost-effective, does not require microscope or incubator. It is recommended as a rapid screening test for the diagnosis of scrub typhus infection in the field or rural area within the hyperendemic region.

Matched MeSH terms: Antibodies, Bacterial/analysis*
Antibody to Pseudomonas pseudomallei exotoxin in sheep exposed to natural infection

Ismail G, Mohamed R, Rohana S, Sharifah HS, Embi N

Vet Microbiol, 1991 May;27(3-4):277-82.
PMID: 1882505

Specific antibody to Pseudomonas pseudomallei exotoxin was detected in sheep sera exposed to natural infection. An enzyme-linked immunosorbent assay (ELISA) was used. Serum antitoxin was present in 49.3% of sera obtained from a flock of sheep naturally exposed to P. pseudomallei infection. Among these sera, 17.0% gave titers of 10,000. In contrast, serum antitoxin was present in only 6.0% of sera collected from sheep kept on a melioidosis-free farm. The ELISA reactivity of all positive sera could be completely absorbed with purified P. pseudomallei exotoxin. Similarly, preincubation of the exotoxin-coated wells with specific antiserum inhibited the ELISA reactivity of sheep sera. The results indicate that exotoxin is produced in vivo during infection by P. pseudomallei.

Matched MeSH terms: Antibodies, Bacterial/blood*
Salmonella agglutinins in normal adult sera in an endemic area

Jamal F, Yasin MS, Abdullah R, Mokhtar N

J Diarrhoeal Dis Res, 1986 Jun;4(2):74-6.
PMID: 3571875
Matched MeSH terms: Antibodies, Bacterial/analysis*
Scrub Typhus antibody in cynomolgus monkeys (Macaca fascicularis) in Malaysia

Heisey GB, Gan E, Shirai A, Groves MG

Lab. Anim. Sci., 1981 Jun;31(3):289-91.
PMID: 6790836

Using an indirect immunofluorescence technique, sera from 113 cynomolgus monkeys (Macaca fascicularis), trapped in Peninsular Malaysia, were screened for the presence of antibody to six prototype strains of Rickettsia tsutsugamushi combined into three polyvalent groupings: I--Karp, TA716, and TA763; II--Gilliam; and III--TA678 and TH1817. Fifteen percent (17/113) of the monkeys had antibody titers greater than or equal to 1:50 to one or more of the antigenic groups. Although a titer greater than or equal to 1:150 is generally considered indicative or prior Rickettsia tsutsugamushi infection, we selected a less than 1:25 titer as a conservative standard to insure non-infected animals. Using this criterion, 62 (55%) of the 113 monkeys were accepted for use in scrub typhus studies. The high prevalence of antibody to scrub typhus in the semi-arboreal cynomolgus monkey is in marked contrast to the low prevalence reported in the strictly arboreal silvered leaf monkeys (Presbytis cristatus). The results of this study indicate that cynomolgus monkeys should be rigorously screened for evidence of prior infection before they are included in experimental scrub typhus studies.

Matched MeSH terms: Antibodies, Bacterial/analysis*
Scrub typhus antibody in mammals in three habitats in Sabah

Muul I, Lim BL, Gan E

Southeast Asian J. Trop. Med. Public Health, 1974 Mar;5(1):80-4.
PMID: 4209140
Matched MeSH terms: Antibodies, Bacterial/analysis*
Ethnic distribution of Chlamydophila pneumoniae antibodies in a Malaysian population and possible correlation with coronary heart disease

Naidu BR, Ngeow YF, Pang T

Eur J Epidemiol, 2003;18(2):135-7.
PMID: 12733835

Chlamydophila pneumoniae, an important respiratory pathogen causing lower respiratory tract infections, has also been implicated in coronary heart disease (CHD). This study reports a cross-sectional, demographic, serological analysis of the prevalence of Chlamydophila pneumoniae antibodies in a multiracial Malaysian population. Generally, Malaysian Indians had the highest degree of seropositivity (58%) followed by the Chinese (54%) and the Malays (32%), results which were statistically significant (CI: 95%; p < 0.01). Interestingly, this trend was also reflected in the study group consisting of patients with acute myocardial infarctions (AMI) and chronic CHD. Again, the Indians were more frequently seropositive (65%), with more than 50% having IgG titres > or = 128. Comparatively, the Chinese and Malays showed 51 and 17% seropositivity respectively. These results were also statistically significant (CI: 95%; p < 0.01). Malaysian Indians are more commonly afflicted with CHD. A variety of factors have been suggested to explain this prevalence including diet, social habits, genetics and the possible role of infectious agents. This study notes an interesting association between this pattern of racial prevalence and the possible role of C. pneumoniae infections as a contributory/predisposing factor in the development of cardiovascular disease.

Matched MeSH terms: Antibodies, Bacterial/immunology*
Fulltext Diagnostic accuracy of rapid diagnostic tests for the early detection of leptospirosis

Alia SN, Joseph N, Philip N, Azhari NN, Garba B, Masri SN, et al.

J Infect Public Health, 2018 11 27;12(2):263-269.
PMID: 30502041 DOI: 10.1016/j.jiph.2018.10.137

BACKGROUND: Leptospirosis is often misdiagnosed with several other tropical febrile illnesses in Malaysia due to similarities in clinical manifestations. Although treatment regimens could be started based on clinical judgments, early diagnosis has become paramount as a guide to chemotherapeutic interventions. Confirmed laboratory diagnosis through MAT or PCR is time consuming and usually available only in reference laboratories and not practical in healthcare settings. Rapid and easy to perform diagnostic tests are widely used in these settings as the point of care diagnosis. The present study was undertaken to compare the diagnostic performance of two IgM based immunodiagnostic assay kits for acute leptospirosis.
METHODS: A total of 50 serum samples were collected from patients clinically suspected for acute leptospirosis on admission in the Hospital Serdang, from June 2016 to June 2017. All the samples were subjected to MAT, lipL32 PCR and the two rapid tests (Leptocheck-WB and ImmuneMed Leptospira IgM Duo Rapid test).
RESULTS: Out of the 50 clinically suspected patients sampled, 19 were confirmed positive for leptospirosis. Six (12%) were confirmed by MAT and 13 (26%) by PCR. Similarly, of the 50 clinically suspected cases, 17 (34%) showed positivity for Leptocheck-WB and 7 (14%) for ImmuneMed Leptospira IgM Duo Rapid test. The overall sensitivity and specificity was 47.37% and 80.65% for Leptocheck-WB, and 21.05% and 90.32% for ImmuneMed Leptospira IgM Duo Rapid test. In another set of previously confirmed MAT positive samples (1:400-1:3600) obtained from a reference laboratory, Leptocheck-WB showed higher sensitivity (90.72%) than ImmuneMed Leptospira IgM Duo Rapid test (40.21%), and comparable specificity for ImmuneMed Leptospira IgM Duo Rapid test (88.89%) and Leptocheck-WB (82.86%).
CONCLUSION: The sensitivity was higher for Leptocheck-WB and had a comparable specificity with ImmuneMed Leptospira IgM Duo Rapid test. Therefore, based on the present study, Leptocheck-WB is found to be a more sensitive rapid immunodiagnostic test for acute leptospirosis screening in hospital settings.

Matched MeSH terms: Antibodies, Bacterial/blood*
Repeated dose toxicity evaluation of a cold chain-free, live, attenuated oral cholera vaccine in Sprague Dawley rats

Xian TH, Parasuraman S, Sinniah K, Ravichandran M, Prabhakaran G

Vaccine, 2019 01 29;37(5):711-720.
PMID: 30630696 DOI: 10.1016/j.vaccine.2018.12.027

The repeated dose toxicity of a prototype cold chain-free, live, attenuated oral cholera vaccine containing 5 × 106 CFU/mL of the VCUSM14P strain was evaluated in Sprague Dawley (SD) rats (single dose administered daily for 30 days) to ascertain its safety for clinical use. Repeated dose toxicity studies for cholera vaccines in the literature have administered 2 or 3 fixed doses at 7, 14, 21 or 69 day intervals. The present study reports an evaluation of 30 repeated doses of cholera vaccine administered at three different concentrations (Group II (1.25 × 106 CFU), Group III (2.5 × 106 CFU) and Group IV (5 × 106 CFU)) in SD rats. The liquid vaccine was administered orally to the rats with the respective dose every day, and normal saline was administered to the control group (Group I). No significant difference (P > 0.05) was observed in the body weights and biochemical parameters of the rats after 15 and 30 repeated doses compared to those of the control group. However, compared to those of Group I, a significant increase (P 

Matched MeSH terms: Antibodies, Bacterial/blood
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: Antibodies, Bacterial/blood
Fulltext Immunogenicity and protective efficacy of a live, oral cholera vaccine formulation stored outside-the-cold-chain for 140 days

Xian TH, Sinniah K, Yean CY, Krishnamoorthy V, Bahari MB, Ravichandran M, et al.

BMC Immunol, 2020 05 25;21(1):29.
PMID: 32450807 DOI: 10.1186/s12865-020-00360-1

BACKGROUND: Cholera, an acute watery diarrhoeal disease caused by Vibrio cholerae serogroup O1 and O139 across the continents. Replacing the existing WHO licensed killed multiple-dose oral cholera vaccines that demand 'cold chain supply' at 2-8 °C with a live, single-dose and cold chain-free vaccine would relieve the significant bottlenecks and cost determinants in cholera vaccination campaigns. In this direction, a prototype cold chain-free live attenuated cholera vaccine formulation (LACV) was developed against the toxigenic wild-type (WT) V. cholerae O139 serogroup. LACV was found stable and retained its viability (5 × 106 CFU/mL), purity and potency at room temperature (25 °C ± 2 °C, and 60% ± 5% relative humidity) for 140 days in contrast to all the existing WHO licensed cold-chain supply (2-8 °C) dependent killed oral cholera vaccines.
RESULTS: The LACV was evaluated for its colonization potential, reactogenicity, immunogenicity and protective efficacy in animal models after its storage at room temperature for 140 days. In suckling mice colonization assay, the LACV recorded the highest recovery of (7.2 × 107 CFU/mL) compared to those of unformulated VCUSM14P (5.6 × 107 CFU/mL) and the WT O139 strain (3.5 × 107 CFU/mL). The LACV showed no reactogenicity even at an inoculation dose of 104-106 CFU/mL in a rabbit ileal loop model. The rabbits vaccinated with the LACV or unformulated VCUSM14P survived a challenge with WT O139 and showed no signs of diarrhoea or death in the reversible intestinal tie adult rabbit diarrhoea (RITARD) model. Vaccinated rabbits recorded a 275-fold increase in anti-CT IgG and a 15-fold increase in anti-CT IgA antibodies compared to those of rabbits vaccinated with unformulated VCUSM14P. Vibriocidal antibodies were increased by 31-fold with the LACV and 14-fold with unformulated VCUSM14P.
CONCLUSION: The vaccine formulation mimics a natural infection, is non-reactogenic and highly immunogenic in vivo and protects animals from lethal wild-type V. cholerae O139 challenge. The single dose LACV formulation was found to be stable at room temperature (25 ± 2 °C) for 140 days and it would result in significant cost savings during mass cholera vaccination campaigns.

Matched MeSH terms: Antibodies, Bacterial/immunology
Evaluation of Salmonella Typhi antigen YncE alongside HlyE for the detection of typhoid fever and its carriers

Franklin F, Chong CW, Chua LH, Anthony AA, Liew MWO, Aziah I, et al.

Med Microbiol Immunol, 2020 Oct;209(5):593-601.
PMID: 32246197 DOI: 10.1007/s00430-020-00667-1

Typhoid fever is a disease caused by Salmonella Typhi that was implicated in millions of illnesses worldwide annually. Individuals that do not recover fully from typhoid fever can become asymptomatic carriers of the disease. Host antibodies against the S. Typhi antigens, HlyE (for acute typhoid) and YncE (for carriers) were previously reported to be useful biomarkers for the disease. Here, we expressed and purified recombinant HlyE and YncE antigens and tested the IgG, IgA and IgM responses in 422 sera samples retrieved from acute typhoid patients, other febrile, food handlers, and healthy individuals. The results showed that HlyE-IgG, -IgA and -IgM ELISAs have a collective sensitivity of 83% while YncE-IgG and -IgA ELISAs identified 16 possible carriers based on their antibody profiles. The identification of sensitive biomarkers for typhoid carrier detection is crucial for disease eradication.

Matched MeSH terms: Antibodies, Bacterial/blood*

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