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  1. Kalai Chelvam K, Yap KP, Chai LC, Thong KL
    PLoS One, 2015;10(5):e0126207.
    PMID: 25946205 DOI: 10.1371/journal.pone.0126207
    Salmonella enterica serovar Typhi (S. Typhi) is a foodborne pathogen that causes typhoid fever and infects only humans. The ability of S. Typhi to survive outside the human host remains unclear, particularly in human carrier strains. In this study, we have investigated the catabolic activity of a human carrier S. Typhi strain in both planktonic and biofilm cells using the high-throughput Biolog Phenotype MicroArray, Minimum Biofilm Eradication Concentration (MBEC) biofilm inoculator (96-well peg lid) and whole genome sequence data. Additional strains of S. Typhi were tested to further validate the variation of catabolism in selected carbon substrates in the different bacterial growth phases. The analyzes of the carbon utilization data indicated that planktonic cells of the carrier strain, S. Typhi CR0044 could utilize a broader range of carbon substrates compared to biofilm cells. Pyruvic acid and succinic acid which are related to energy metabolism were actively catabolised in the planktonic stage compared to biofilm stage. On the other hand, glycerol, L-fucose, L-rhamnose (carbohydrates) and D-threonine (amino acid) were more actively catabolised by biofilm cells compared to planktonic cells. Notably, dextrin and pectin could induce strong biofilm formation in the human carrier strain of S. Typhi. However, pectin could not induce formation of biofilm in the other S. Typhi strains. Phenome data showed the utilization of certain carbon substrates which was supported by the presence of the catabolism-associated genes in S. Typhi CR0044. In conclusion, the findings showed the differential carbon utilization between planktonic and biofilm cells of a S. Typhi human carrier strain. The differences found in the carbon utilization profiles suggested that S. Typhi uses substrates mainly found in the human biliary mucus glycoprotein, gallbladder, liver and cortex of the kidney of the human host. The observed diversity in the carbon catabolism profiles among different S. Typhi strains has suggested the possible involvement of various metabolic pathways that might be related to the virulence and pathogenesis of this host-restricted human pathogen. The data serve as a caveat for future in-vivo studies to investigate the carbon metabolic activity to the pathogenesis of S. Typhi.
    Matched MeSH terms: Typhoid Fever/microbiology
  2. Choo KE, Razif AR, Oppenheimer SJ, Ariffin WA, Lau J, Abraham T
    J Paediatr Child Health, 1993 Feb;29(1):36-9.
    PMID: 8461177
    Data are presented for 2382 children investigated for fever in a Malaysian hospital between 1984 and 1987 when Widal tests and blood cultures were a routine part of every fever screen. There were 145 children who were culture positive (TYP-CP) for Salmonella typhi, while 166 were culture negative but were diagnosed as having typhoid (TYP-CN). Analyses of the sensitivity and specificity of combinations of initial Widal titres in predicting a positive S. typhi culture in a febrile child (culture positive vs the rest) showed the best model to be an O- and/or H-titre of > or = 1 in 40 (sensitivity 89%; specificity 89%). While the negative predictive value of the model was high (99.2%) the positive predictive value remained below 50% even for very high titres of O and H (> 1 in 640), at which point the specificity was 98.5%, supporting the clinical view that a high proportion of the TYP-CN patients really were typhoid but were missed by culture. The TYP-CN patients showed a very similar clinical and age profile to TYP-CP patients. The length of history of fever did not affect the initial Widal titre in culture positive cases. The Widal test in children remains a sensitive and specific 'fever screen' for typhoid although it will not identify all cases. In children, lower cut-off points for O- and H-titres should be used than are generally recommended.
    Matched MeSH terms: Typhoid Fever/microbiology
  3. Pang T, Levine MM, Ivanoff B, Wain J, Finlay BB
    Trends Microbiol., 1998 Apr;6(4):131-3.
    PMID: 9587187
    Matched MeSH terms: Typhoid Fever/microbiology
  4. Muhammad EN, Abdul Mutalip MH, Hasim MH, Paiwai F, Pan S, Mahmud MAF, et al.
    BMC Infect Dis, 2020 Nov 16;20(1):843.
    PMID: 33198646 DOI: 10.1186/s12879-020-05500-x
    BACKGROUND: Typhoid fever causes global morbidity and mortality and is a significant health burden, particularly in low- and middle-income countries. The direct fecal-oral route is the main transmission mode, but indirect environmental transmission could occur, particularly in urban settings. This study aimed to investigate the burden and trend of typhoid fever, reporting the coverage system between government and private practice and pattern of multidrug-resistant (MDR) typhoid cases in the urban Klang Valley area from 2011 to 2015.

    METHODS: The data from a cross-sectional study retrieved from the e-Notifikasi System, a national reporting system for communicable diseases provided by the Disease Control Division, Ministry of Health Malaysia and secondary data of all the typhoid cases were obtained from the public and private hospitals and laboratories in Klang Valley. Descriptive analysis was performed to examine the sociodemographic characteristics, spatial mapping was conducted to examine trends, and the crude incidence rates of confirmed typhoid cases and percentage of reporting coverage were calculated. Significant differences between MDR and non-MDR Salmonella typhi were determined in the patient's sociodemographic characteristics, which were analyzed using χ2 test. P values typhoid fever cases were reported in Klang Valley; however, only 265 cases were confirmed by culture tests. The crude incidence rates of confirmed cases were between 0.5 to 0.7 but peaked at 1.42 per 100,000 population in 2015. Most typhoid fever cases were observed among men (55.6%), individuals aged 21 to 30 years (27.6%), Malaysians (86.3%) and individuals of Malay ethnicity (52.1%). The reporting coverage of confirmed cases was 78.9% and non-reporting coverage of unconfirmed typhoid cases was 79.5%. The predictive value positive (PVP) was 89.3, and 7.5% were detected as MDR Salmonella typhi. Statistical significance was found in gender, citizenship and ethnicity regarding MDR Salmonella typhi (p = 0.004, p = 0.008 and p = 0.034, respectively).

    CONCLUSIONS: The local transmission of typhoid is still prevalent in the Klang Valley despite rapid urbanization and development in recent years. These findings are essential for policy makers to plan and implement focused and effective preventative activities to curb typhoid infection in urban areas.

    Matched MeSH terms: Typhoid Fever/microbiology
  5. Yap KP, Thong KL
    Trop Med Int Health, 2017 08;22(8):918-925.
    PMID: 28544285 DOI: 10.1111/tmi.12899
    Next-generation whole-genome sequencing has revolutionised the study of infectious diseases in recent years. The availability of genome sequences and its understanding have transformed the field of molecular microbiology, epidemiology, infection treatments and vaccine developments. We review the key findings of the publicly accessible genomes of Salmonella enterica serovar Typhi since the first complete genome to the most recent release of thousands of Salmonella Typhi genomes, which remarkably shape the genomic research of S. Typhi and other pathogens. Important new insights acquired from the genome sequencing of S. Typhi, pertaining to genomic variations, evolution, population structure, antibiotic resistance, virulence, pathogenesis, disease surveillance/investigation and disease control are discussed. As the numbers of sequenced genomes are increasing at an unprecedented rate, fine variations in the gene pool of S. Typhi are captured in high resolution, allowing deeper understanding of the pathogen's evolutionary trends and its pathogenesis, paving the way to bringing us closer to eradication of typhoid through effective vaccine/treatment development.
    Matched MeSH terms: Typhoid Fever/microbiology*
  6. Combs BG, Passey M, Michael A, Pang T, Lightfoot D, Alpers MP
    P N G Med J, 2005 Sep-Dec;48(3-4):158-67.
    PMID: 17212062
    The prevalence of typhoid in the Papua New Guinea (PNG) highlands region increased rapidly in the mid-1980s, and now remains endemic. In this study ribotyping has been used to examine the number and types of Salmonella enterica serovar Typhi strains present during the 1977-1996 period. The ribotyping banding pattern results were based on Cla I and Eco RV digests. The 57 PNG isolates were divided into 11 different ribotypes. Comparison of ribotypes using coefficient of similarity values revealed a diverse group of ribotypes. Several strains appear to be endemic in PNG For instance, ribotypes 1, 2 and 3 were most commonly found among PNG isolates and isolates with these ribotypes have been cultured over a period of at least 11 years (1985-1996). Ribotype 3 was also observed in isolates from Malaysia and Thailand. Also found in PNG were ribotypes 4, 5, 6, 7, 8, 9, 16 and 17. The ribotyping suggests that serovar Typhi strains present in PNG include unique strains of serovar Typhi and also strains that are common to other countries.
    Matched MeSH terms: Typhoid Fever/microbiology
  7. Abdullah J, Saffie N, Sjasri FA, Husin A, Abdul-Rahman Z, Ismail A, et al.
    Braz J Microbiol, 2014;45(4):1385-91.
    PMID: 25763045
    An in-house loop-mediated isothermal amplification (LAMP) reaction was established and evaluated for sensitivity and specificity in detecting the presence of Salmonella Typhi (S. Typhi) isolates from Kelantan, Malaysia. Three sets of primers consisting of two outer and 4 inner were designed based on locus STBHUCCB_38510 of chaperone PapD of S. Typhi genes. The reaction was optimised using genomic DNA of S. Typhi ATCC7251 as the template. The products were visualised directly by colour changes of the reaction. Positive results were indicated by green fluorescence and negative by orange colour. The test was further evaluated for specificity, sensitivity and application on field samples. The results were compared with those obtained by gold standard culture method and Polymerase Chain Reaction (PCR). This method was highly specific and -10 times more sensitive in detecting S. Typhi compared to the optimised conventional polymerase chain reaction (PCR) method.
    Matched MeSH terms: Typhoid Fever/microbiology
  8. Koay AS, Jegathesan M, Rohani MY, Cheong YM
    PMID: 9322288
    Strains of Salmonella typhi implicated in two separate cases of laboratory acquired infection from patients and the medical laboratory technologists who processed the patients' samples were analysed by pulsed-field gel electrophoresis. Although all four isolates were of bacteriophage type E1, PFGE was able to demonstrate that the strains responsible for the two laboratory acquired cases were not genetically related. The PFGE patterns of the isolates from the MLTs were found to be identical to those of the corresponding patients after digestion with restriction enzyme AvrII. This provided genetic as well as epidemiological evidence for the source of the laboratory acquired infections.
    Matched MeSH terms: Typhoid Fever/microbiology
  9. Jegathesan M
    J Hyg (Lond), 1983 Feb;90(1):91-7.
    PMID: 6822730
    The pattern of phage types of 2553 strains of Salmonella typhi isolated over the 10-year period 1970-9 was studied. During the period 29 different phage types were encountered, not including the categories of 'untypable strains', 'degraded Vi-strains' and Vi negative strains. For the period as a whole, the commonest phage types encountered were A (20.9%), E1 (14.8%), D1 (10.3%), degraded Vi positive strains (10.3%), untypable Vi strains (7.3%), C4 (7.1%), D2 (4.4%), E2 (3.9%) and type 25 (2.6%). There were phage types which appeared in the early years of the period and then disappeared (types B2, D9 and D1-N). Others only made their appearance in recent years (K1 and 53). Notable differences were also seen in the predilection of some phage types for certain geographical areas.
    Matched MeSH terms: Typhoid Fever/microbiology*
  10. Chin CF, Teh BA, Anthony AA, Aziah I, Ismail A, Ong EB, et al.
    Appl Biochem Biotechnol, 2014 Nov;174(5):1897-906.
    PMID: 25149461 DOI: 10.1007/s12010-014-1173-y
    In our earlier study, an immunoblot analysis using sera from febrile patients revealed that a 50-kDa band from an outer membrane protein fraction of Salmonella enterica serovar Typhi was specifically recognized only by typhoid sera and not sera from other febrile illnesses. Here, we investigated the identities of the proteins contained in the immunogenic 50-kDa band to pinpoint antigens responsible for its immunogenicity. We first used LC-MS/MS for protein identification, then used the online tool ANTIGENpro for antigenicity prediction and produced recombinant proteins of the lead antigens for validation in an enzyme-linked immunosorbent assay (ELISA). We found that proteins TolC, GlpK and SucB were specific to typhoid sera but react to antibodies differently under native and denatured conditions. This difference suggests the presence of linear and conformational epitopes on these proteins.
    Matched MeSH terms: Typhoid Fever/microbiology
  11. Thong KL, Bhutta ZA, Pang T
    Int J Infect Dis, 2000;4(4):194-7.
    PMID: 11231181
    OBJECTIVE: The goal of this study was to report the molecular analysis of antibiotic-sensitive and multidrug-resistant (MDR) strains of Salmonella typhi, using pulsed-field gel electrophoresis (PFGE), with a particular emphasis on the coexistence of these strains in a typhoid-endemic region of Karachi, Pakistan.

    METHODS: One hundred isolates of S. typhi in humans (50 MDR and 50 antibiotic-sensitive isolates) from sporadic cases of typhoid fever were analyzed by Vi-phage typing, antibiograms and PFGE.

    RESULTS: The MDR S. typhi strains were resistant to ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole. Analysis by PFGE showed that 50 MDR isolates of S. typhi had a single, homogenous PFGE profile, which was distinctly different from that of 50 antibiotic-sensitive isolates obtained in the same time frame from the same area. This latter group of isolates showed much greater diversity of PFGE profiles, as has been observed in other endemic regions.

    CONCLUSIONS: Multidrug-resistant and antibiotic-susceptible strains of S. typhi can coexist in endemic areas as epidemiologically independent pathogens and are not in competition for continued persistence and transmission.

    Matched MeSH terms: Typhoid Fever/microbiology*
  12. Thong KL, Passey M, Clegg A, Combs BG, Yassin RM, Pang T
    J Clin Microbiol, 1996 Apr;34(4):1029-33.
    PMID: 8815078
    Molecular characterization of a total of 52 human isolates of Salmonella typhi from Papua New Guinea was performed by using pulsed-field gel electrophoresis (PFGE) after digestion of chromosomal DNA with three restriction endonucleases, XbaI (5'-TCTAGA-3'), AvrII (5'-CCTAGG-3'), and SpeI (5'-ACTAGT-3'). Of the 52 isolates tested, 11 were obtained from patients with fatal typhoid fever and 41 were obtained from patients with nonfatal disease. The 52 isolates showed limited genetic diversity as evidenced by only three different PFGE patterns detected following digestion with XbaI (patterns X1 to X3; F [coefficient of similarity] = 0.86 to 1.0), four patterns detected following digestion with AvrII (patterns A1 to A4; F =0.78 to 1.0), and two patterns detected following digestion with SpeI (patterns S1 and S2; F = 0.97 to 1.0). Of the 52 isolates, 37 were phage typed, and all belonged to phage type D2. All 11 isolates obtained from patients with fatal typhoid fever were identical (F = 1.0) and possessed the PFGE pattern combination X1S1A1, whereas the 41 isolates from patients with nonfatal typhoid fever had various PFGE pattern combinations, the most common being X2S1A2 (39%), X1S1A1 (24%), and X1S1A2 (15%). Thus, all the isolates from patients with the fatal disease had the X1 and A1 patterns, whereas the majority of the isolates from patients with nonfatal typhoid fever possessed the X2 and A2 patterns. The data suggest that there is an association among strains of S. typhi between genotype, as assessed by PFGE patterns, and the capability to cause fatal illness. Analysis of blood and fecal isolates of S. typhi from the same patient also indicated that some genetic changes occur in vivo during the course of infection.
    Matched MeSH terms: Typhoid Fever/microbiology*
  13. Thong KL, Cordano AM, Yassin RM, Pang T
    Appl Environ Microbiol, 1996 Jan;62(1):271-4.
    PMID: 8572705
    Molecular characterization of a total of 54 isolates of Salmonella typhi from Santiago, Chile, was performed by pulsed-field gel electrophoresis (PFGE) after digestion of chromosomal DNA with three restriction endonucleases: XbaI (5'-TCTAGA-3'), AvrII (5'-CCTAGG-3'), and SpeI (5'-ACTAGT-3'). Thirteen of the 54 isolates were obtained from environmental sources (sewage and river water), and the rest were isolates from clinical cases of typhoid fever. Considerable genetic diversity was detected among the human isolates obtained in 1994, as evidenced by the presence of 14 to 19 different PFGE patterns among 20 human isolates, with F (coefficient of similarity) values ranging from 0.69 to 1.0 (XbaI), 0.61 to 1.0 (AvrII), and 0.70 to 1.0 (SpeI). A total of eight phage types were detected among these 20 isolates, with 50% possessing the E1 or 46 phage type. There was no correlation between PFGE pattern and phage types. Similar diversity was seen among 21 isolates obtained in 1983, with 17 to 19 PFGE patterns detected and F values of 0.56 to 1.0 (XbaI), 0.55 to 1.0 (AvrII), and 0.67 to 1.0 (SpeI). Comparison of these two groups of human isolates obtained 11 years apart indicated that certain molecular types of S. typhi are shared and are able to persist for considerable periods. A similar degree of genetic diversity was also detected among the environmental isolates of S. typhi, for which 10 to 12 different PFGE patterns were detected among the 13 isolates analyzed, with F values ranging from 0.56 to 1.0 (XbaI), 0.52 to 1.0 (AvrII), and 0.69 to 1.0 (SpeI). Certain molecular types present among the environmental isolates of S. typhi were also found among the human isolates from the same time period, providing evidence for the epidemiological link between environmental reservoirs and human infection.
    Matched MeSH terms: Typhoid Fever/microbiology
  14. Yap KP, Teh CS, Baddam R, Chai LC, Kumar N, Avasthi TS, et al.
    J Bacteriol, 2012 Sep;194(18):5124-5.
    PMID: 22933756 DOI: 10.1128/JB.01062-12
    Salmonella enterica serovar Typhi is the causative agent of typhoid fever, which causes nearly 21.7 million illnesses and 217,000 deaths globally. Herein, we describe the whole-genome sequence of the Salmonella Typhi strain ST0208, isolated from a sporadic case of typhoid fever in Kuala Lumpur, Malaysia. The whole-genome sequence and comparative genomics allow an in-depth understanding of the genetic diversity, and its link to pathogenicity and evolutionary dynamics, of this highly clonal pathogen that is endemic to Malaysia.
    Matched MeSH terms: Typhoid Fever/microbiology
  15. Thong KL, Goh YL, Yasin RM, Lau MG, Passey M, Winston G, et al.
    J Clin Microbiol, 2002 Nov;40(11):4156-60.
    PMID: 12409390
    Pulsed-field gel electrophoresis (PFGE) of XbaI-digested chromosomal DNA was performed on 133 strains of Salmonella enterica serovar Typhi obtained from Papua New Guinea, with the objective of assessing the temporal variation of these strains. Fifty-two strains that were isolated in 1992 and 1994 were of one phage type, D2, and only two predominant PFGE profiles, X1 and X2, were present. Another 81 strains isolated between 1997 and 1999 have shown divergence, with four new phage types, UVS I (n = 63), UVS (n = 5), VNS (n = 4), and D1 (n = 9), and more genetic variability as evidenced by the multiple and new PFGE XbaI profiles (21 profiles; Dice coefficient, F = 0.71 to 0.97). The two profiles X1 and X2 have remained the stable, dominant subtypes since 1992. Cluster analysis based on the unweighted pair group method using arithmetic averages algorithm identifies two main clusters (at 87% similarity), indicating that the divergence of the PFGE subtypes was probably derived from some genomic mutations of the X1 and X2 subtypes. The majority of isolates were from patients with mild and moderate typhoid fever and had various XbaI profiles. A single isolate from a patient with fatal typhoid fever had a unique X11 profile, while four of six isolates from patients with severe typhoid fever had the X1 pattern. In addition, 12 paired serovar Typhi isolates recovered from the blood and fecal swabs of individual patients exhibited similar PFGE patterns, while in another 11 individuals paired isolates exhibited different PFGE patterns. Three pairs of isolates recovered from three individuals had different phage types and PFGE patterns, indicating infection with multiple strains. The study reiterates the usefulness of PFGE in assessing the genetic diversity of S. enterica serovar Typhi for both long-term epidemiology and in vivo stability and instability within an individual patient.
    Matched MeSH terms: Typhoid Fever/microbiology
  16. Cheong YM, Jegathesan M
    Med J Malaysia, 1992 Dec;47(4):331.
    PMID: 1303490
    Matched MeSH terms: Typhoid Fever/microbiology
  17. Goay YX, Chin KL, Tan CL, Yeoh CY, Ja'afar JN, Zaidah AR, et al.
    Biomed Res Int, 2016;2016:8905675.
    PMID: 27975062
    Salmonella Typhi (S. Typhi) causes typhoid fever which is a disease characterised by high mortality and morbidity worldwide. In order to curtail the transmission of this highly infectious disease, identification of new markers that can detect the pathogen is needed for development of sensitive and specific diagnostic tests. In this study, genomic comparison of S. Typhi with other enteric pathogens was performed, and 6 S. Typhi genes, that is, STY0201, STY0307, STY0322, STY0326, STY2020, and STY2021, were found to be specific in silico. Six PCR assays each targeting a unique gene were developed to test the specificity of these genes in vitro. The diagnostic sensitivities and specificities of each assay were determined using 39 S. Typhi, 62 non-Typhi Salmonella, and 10 non-Salmonella clinical isolates. The results showed that 5 of these genes, that is, STY0307, STY0322, STY0326, STY2020, and STY2021, demonstrated 100% sensitivity (39/39) and 100% specificity (0/72). The detection limit of the 5 PCR assays was 32 pg for STY0322, 6.4 pg for STY0326, STY2020, and STY2021, and 1.28 pg for STY0307. In conclusion, 5 PCR assays using STY0307, STY0322, STY0326, STY2020, and STY2021 were developed and found to be highly specific at single-gene target resolution for diagnosis of typhoid fever.
    Matched MeSH terms: Typhoid Fever/microbiology*
  18. Thong KL, Puthucheary SD, Pang T
    Res. Microbiol., 1997 Mar-Apr;148(3):229-35.
    PMID: 9765803
    We performed genome size estimation of 17 recent human isolates of Salmonella typhi from geographically diverse regions using pulsed-field gel electrophoresis (PFGE) after digestion of chromosomal DNA with restriction endonucleases XbaI (5'-TCTAGA-3'), AvrII (5'-CCTAGG-3') and SpeI (5'-ACTAGT-3'), and summation of the sizes of restriction fragments obtained. All 17 isolates had circular chromosomes, and genome sizes differed by as much as 959 kb, ranging from 3,964 to 4,923 kb (mean genome size = 4,528 kb). The data obtained confirm the usefulness of PFGE in studies of bacterial genome size and are in agreement with recent results indicating considerable genetic diversity and genomic plasticity of S. typhi. The variation in genome sizes noted may be relevant to the observed biological properties of this important human pathogen, including its virulence.
    Matched MeSH terms: Typhoid Fever/microbiology*
  19. Yap KP, Gan HM, Teh CS, Baddam R, Chai LC, Kumar N, et al.
    J Bacteriol, 2012 Nov;194(21):5970-1.
    PMID: 23045488 DOI: 10.1128/JB.01416-12
    Salmonella enterica serovar Typhi is a human pathogen that causes typhoid fever predominantly in developing countries. In this article, we describe the whole genome sequence of the S. Typhi strain CR0044 isolated from a typhoid fever carrier in Kelantan, Malaysia. These data will further enhance the understanding of its host persistence and adaptive mechanism.
    Matched MeSH terms: Typhoid Fever/microbiology
  20. Pang T, Altwegg M, Martinetti G, Koh CL, Puthucheary S
    Microbiol. Immunol., 1992;36(5):539-43.
    PMID: 1513268
    Genetic variation among Malaysian isolates of Salmonella typhi was determined by analysis of ribosomal RNA gene restriction patterns. Of the 20 isolates analyzed, eight different pattern combinations were detected. The amount of variation observed was also dependent upon the restriction endonuclease used; PstI produced more different patterns than did SmaI. The results suggested that disease activity was due to a number of different clones circulating simultaneously rather than a single strain. Further implications of the data are discussed.
    Matched MeSH terms: Typhoid Fever/microbiology
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