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  1. Fulltext First report of pathogenic Leptospira spp. isolated from urine and kidneys of naturally infected cats
    Alashraf AR, Lau SF, Khairani-Bejo S, Khor KH, Ajat M, Radzi R, et al.
    PLoS One, 2020;15(3):e0230048.
    PMID: 32155209 DOI: 10.1371/journal.pone.0230048
    Leptospirosis is one of the most widespread zoonotic diseases and can infect both humans and animals worldwide. Healthy cat, as a potential source of exposure to humans, are likely underestimated owing to the lack of overt clinical signs associated with Leptospira spp. infection in this species. The aim of the study was to determine the exposure, shedding, and carrier status of leptospires in shelter cats in Malaysia by using serological, molecular, and bacteriological methods. For this study, 82 healthy cats from two shelters were sampled. The blood, urine, and kidneys were tested using the microscopic agglutination test (MAT), polymerase chain reaction (PCR), and bacterial culture. On the basis of serological, molecular, and/or culture techniques, the total detection of leptospiral infection was 29.3% (n = 24/82). Through culture techniques, 16.7% (n = 4/24) of the cats that tested positive were carriers with positive kidney cultures, and one cat was culture positive for both urine and kidney. The Leptospira spp. isolates were identified as pathogenic L. interrogans serovar Bataviae through serological and molecular methods. Through serological techniques, 87.5% (n = 21/24) had positive antibody titers (100-1600) and most of the Bataviae serogroup (n = 19/21). Using PCR, 16.7% (n = 4/24) of cats were shown to have pathogenic Leptospira spp. DNA in their urine. Furthermore, three out of four culture positive cats were serology negative. The present study reports the first retrieval of pathogenic leptospires from urine and kidneys obtained from naturally infected cats. The results provide evidence of the potential role of naturally infected cats in the transmission of leptospires. Additionally, leptospiral infection occurs sub-clinically in cats. The culture isolation provides evidence that healthy cats could be reservoirs of leptospiral infection, and this information may promote the development of disease prevention strategies for the cat population.
  2. Fulltext A case-control study of risk factors for bovine brucellosis seropositivity in Peninsular Malaysia
    Anka MS, Hassan L, Khairani-Bejo S, Zainal MA, Mohamad RB, Salleh A, et al.
    PLoS One, 2014;9(9):e108673.
    PMID: 25265020 DOI: 10.1371/journal.pone.0108673
    Bovine brucellosis was first reported in Peninsular Malaysia in 1950. A subsequent survey conducted in the country revealed that the disease was widespread. Current knowledge on the potential risk factors for brucellosis occurrence on cattle farms in Malaysia is lacking. Therefore, we conducted a case-control study to identify the potential herd-level risk factors for bovine brucellosis occurrence in four states in the country, namely Kelantan, Pahang, Selangor and Negeri Sembilan. Thirty-five cases and 36 controls of herds were selected where data on farm management, biosecurity, medical history and public health were collected. Multivariable logistic regression identified that Brucella seropositive herds were more likely to; have some interaction with wildlife (OR 8.9, 95% CI = 1.59-50.05); originated from farms where multiple species such as buffalo/others (OR 41.8, 95% CI = 3.94-443.19) and goat/sheep (OR 8.9, 95%Cl = 1.10-71.83) were reared, practice extensive production system (OR 13.6, 95% CI 1.31-140.24) and have had episodes of abortion in the past (OR 51.8, 95% CI = 4.54-590.90) when compared to seronegative herds. Considering the lack of information on the epidemiology of bovine brucellosis in peninsular Malaysia and absence of information on preventing the inception or spread of the disease, this report could contribute to the on-going area-wise national brucellosis eradication program.
  3. Staphylococcal cassette chromosome mec (SCCmec) and characterization of the attachment site (attB) of methicillin resistant Staphylococcus aureus (MRSA) and methicillin susceptible Staphylococcus aureus (MSSA) isolates
    Bitrus AA, Zunita Z, Khairani-Bejo S, Othman S, Ahmad Nadzir NA
    Microb Pathog, 2018 Oct;123:323-329.
    PMID: 30053600 DOI: 10.1016/j.micpath.2018.07.033
    This study was designed to screen for SCCmec types and to characterize the attachment site (attB) and universal insertion site (orfX) of SCCmec in a collection of 27 isolates (n = 11) methicillin resistant S. aureus and (n = 16) methicillin susceptible S. aureus isolates in Malaysia. Screening of SCCmec types and characterization of the attachment site was carried out using PCR amplification and Sanger's sequencing method. The result showed that a large proportion of the MRSA isolates carried SCCmec type III 7/11 (63%). Three isolates 3/11 (27%) and 1/11 (9.0%) carried SCCmec type II and IVd respectively. Amplification of the universal insertion site of the SCCmec (orfX) and attachment site (attB) showed that all 16 S. aureus isolates were positive for the orfX gene, while only 7 were positive for the attB gene. Phylogenetic diversity showed that the isolates clustered around strains with features similar to a community acquired MRSA. In conclusion, a high carriage rate of SCCmec type III was observed. The result also showed that all the S. aureus isolates have the orfX structure; however, not all isolates possesses the attB site on the 3' end of the orfX region.
  4. Fulltext Seroprevalence and Distribution of Leptospiral Serovars in Livestock (cattle, Goats, and Sheep) in Flood-prone Kelantan, Malaysia
    Sabri Abdul Rahman M, Khairani Bejo S, Zakaria Z, Hassan L, Azri Roslan M
    J Vet Res, 2021 Mar;65(1):53-58.
    PMID: 33817395 DOI: 10.2478/jvetres-2021-0003
    Introduction: Leptospirosis is a bacterial disease that affects both humans and animals, the occurrence of which increases markedly during and after heavy rainfall and flooding. The aim of this study was to determine the serological prevalence of leptospiral infection in livestock after a voluminous flood in 10 districts of the Malaysian state of Kelantan.

    Material and Methods: In December 2014, Kelantan was hit by an extensive flood. A total of 1,728 serum samples were collected from livestock from the state, comprised of 1,024 from cattle, 366 from goats and 338 from sheep, and they were tested using the microscopic agglutination test (MAT).

    Results: Altogether, 203 (11.75%; 203/1728; 95% CI: 10.20%-13.30%) of the tested sera were found to be positive serologically. Cattle had the highest prevalence of 14.16% (145/1024), while goats and sheep had 11.20% (41/366) and 5.03% (17/338) respectively. The most frequent serovars detected were Hardjo-bovis (3.70%; 64/1728), Hebdomadis (2.08%; 36/1728) and Pomona (1.04%; 18/1728). There was a statistically significant association (P < 0.05) between livestock that were exposed to the flood and seropositivity.

    Conclusion: This study showed that flood is a risk factor that can play a role in the epidemiology of leptospiral infection in livestock.

  5. Fulltext Rapid detection of pathogenic leptospires by lyophilized reagent-based polymerase chain reaction
    Lee SV, Tai ES, Mutalib AR, Khairani-Bejo S, Bahaman AR
    Trop Biomed, 2011 Dec;28(3):497-505.
    PMID: 22433877 MyJurnal
    A simple and reliable tool for the early diagnosis of leptospirosis is urgently needed. We report the development of a lyophilized reagent-based polymerase chain reaction (PCR) assay targeting lipL32 gene, which is present only in pathogenic leptospires. To determine the effectiveness of the newly developed assay in the early diagnosis of leptospirosis, the sensitivity and specificity was evaluated. In simulated clinical samples, the assay was able to detect 10² and 10³ leptospires/ml in spiked urine and blood samples, respectively. In experimentally infected animals, leptospiral DNA could be detected in blood and lung samples as early as Day 1 post infection. This assay was also shown to be stable and remained sensitive for up to five months at ambient temperature. Hence, this lyophilized reagent-based PCR assay with high specificity, sensitivity and stability would provide a simple, rapid and reliable method in diagnosing acute leptospirosis, especially in the field of veterinary medicine.
  6. Detection and characterization of Leptospira spp. in dogs diagnosed with kidney and/or liver disease in Selangor, Malaysia
    Rahman SA, Khor KH, Khairani-Bejo S, Lau SF, Mazlan M, Roslan A, et al.
    J Vet Diagn Invest, 2021 Sep;33(5):834-843.
    PMID: 34148436 DOI: 10.1177/10406387211024575
    Leptospirosis is a serious bacterial disease that affects both humans and animals. A wide range of symptoms have been described in humans; the disease in dogs is commonly associated with kidney and/or liver disease. In Malaysia, information about the common serovars infecting dogs is limited. Therefore, we investigated the occurrences of leptospirosis in 124 pet dogs diagnosed with kidney and/or liver disease. Blood, urine, abdominal effusion, and/or kidney and liver were collected from the dogs. Based on microscopic agglutination testing, 53 of 124 (42.7%) dogs were seropositive for leptospiral exposure. Sera were frequently positive to serovars Bataviae (n = 12), Javanica (n = 10), and Icterohaemorrhagiae (n = 10). Direct detection using PCR showed that 42 of 124 (33.9%) of the whole blood and 36 of 113 (31.9%) urine samples were positive for pathogenic Leptospira spp. By PCR, 2 of 23 (9.1%) kidney and 2 of 23 (9.1%) liver were positive for pathogenic Leptospira spp. Abdominal effusion from 4 dogs were PCR-positive for pathogenic Leptospira spp. The species detected were L. interrogans, L. borgpetersenii, L. kirschneri, and L. kmetyi by partial 16S rRNA sequencing. We further identified and characterized 11 Leptospira spp. isolates from 8 dogs as serovars Bataviae, Javanica, and Australis. The mortality rate of the Leptospira-infected dogs was high (18 of 53; 34%).
  7. The prevalence and distribution of Brucella melitensis in goats in Malaysia from 2000 to 2009
    Bamaiyi PH, Hassan L, Khairani-Bejo S, ZainalAbidin M, Ramlan M, Adzhar A, et al.
    Prev Vet Med, 2015 May 1;119(3-4):232-6.
    PMID: 25746928 DOI: 10.1016/j.prevetmed.2015.02.001
    A study was conducted to describe the prevalence and distribution of zoonotic Brucella melitensis in goats in Peninsular Malaysia. Using serosurveillance data of the last decade (2000-2009) involving 119,799 goats and 3555 farms, the seroprevalence of brucellosis among goats was 0.91% (95% CI=0.86-0.96) and among farms was 7.09% (95% CI=6.27-7.98). The odds of brucellosis was significantly (P<0.05) higher in the later part of the decade, in larger herd size and among the states located in the peninsula as compared to eastern Malaysia. The infection was detected throughout Malaysia but at generally low seroprevalences with states like Perlis that border neighbouring countries having higher seroprevalence of brucellosis than other non-border states.
  8. Case-control study on risk factors associated with Brucella melitensis in goat farms in Peninsular Malaysia
    Bamaiyi PH, Hassan L, Khairani-Bejo S, ZainalAbidin M, Ramlan M, Krishnan N, et al.
    Trop Anim Health Prod, 2014 Jun;46(5):739-45.
    PMID: 24633657 DOI: 10.1007/s11250-014-0557-x
    Caprine brucellosis is a bacterial zoonotic infection affecting goats especially in developing countries all over the world. In Malaysia, the risk factors associated with this infection in farms have not been studied. A case-control study was carried out in goat farms in four states of Malaysia to elucidate the risk factors associated with the infection on the farms using structured questionnaires and face-to-face interviews. Results indicate that the introduction of new animals (OR = 5.25; 90 % CI = 1.46, 18.88), younger age category of farms (OR = 5.53; 90 % CI = 1.09, 21.66), and farms with single breed of goats (OR = 8.50; 90 % CI = 1.27, 41.97) were significant risk factors for brucellosis. In order to control brucellosis or possibly eradicate it in goat farms, these factors need to be dealt with. Enforcing stringent importation protocols or complete ban of goat importation from brucellosis endemic countries will help reduce risk of introducing new infection into the country.
  9. Fulltext Isolation and molecular characterization of Brucella melitensis from seropositive goats in Peninsula Malaysia
    Bamaiyi PH, Hassan L, Khairani-Bejo S, Zainal Abidin M, Ramlan M, Krishnan N, et al.
    Trop Biomed, 2012 Dec;29(4):513-8.
    PMID: 23202595
    A study was carried out to isolate Brucella melitensis using established bacteriological and PCR techniques in Brucella seropositive goats in farms in Selangor, Negeri Sembilan, Melaka and Pulau Pinang. Brucella melitensis was isolated from 7 of 134 reactors with the highest isolation from the vaginal swabs (57.14%) followed by the spleen (28.57%), uterine fluid (14.29%). No Brucella was isolated from the lymph nodes. PCR confirmed all the seven isolates as B. melitensis and isolates were phylogenetically related to other isolates from India, Iran, and Israel but most closely related to isolates from Singapore.
  10. Fulltext Development of indirect IgG elisa for detection of leptospiral infection in cattle using Malaysian local isolate Leptospira kmetyi serovar Malaysia strain Bejo-Iso9
    Sabri, A.R., Khairani-Bejo, S., Zunita, Z., Hassan, L.
    Jurnal Veterinar Malaysia, 2017;29(2):5-8.
    MyJurnal
    Cattle is one of an important maintenance hosts involved in the transmission of leptospirosis. Serological method is always implemented to detect current or past leptospiral infection in cattle. The results can be obtained immediately compared to isolation method which need more longer time. An in-house IgG enzyme-linked immunosorbent assay (ELISA) using heat-killed whole cells of local isolate Leptospira kmetyi serovar malaysia strain Bejo-Iso9 (ELISABejo-Iso9) was developed in this study. The perfomance of ELISA-Bejo-Iso9 was compared with in-house IgGELISA using reference strain 117123 (hardjo bovis) antigen (ELISA-117123) and CUSABIO® commercial ELISA. It was found that, the performance of ELISA-Bejo-Iso9 was promising to compete with commercial ELISA. The specificity and sensitivity was 98.75% and 53.75%, respectively. The sensitivity of ELISA-Bejo-Iso9 was lower than commercial ELISA. However, the specificity of ELISA-Bejo-Iso9 was higher than commercial ELISA. Due to economically and availability factors, this finding suggested that the ELISA-Bejo-Iso9 can be used as an alternatif method for serological diagnosis of leptospirosis in cattle.
  11. Fulltext Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against leptospira for early detection of acute leptospirosis
    Seenichamy A, Bahaman AR, Mutalib AR, Khairani-Bejo S
    Biomed Res Int, 2014;2014:592858.
    PMID: 24860824 DOI: 10.1155/2014/592858
    Leptospirosis is one of the zoonotic diseases in animals and humans throughout the world. LipL21 is one of the important surface-exposed lipoproteins in leptospires and the most effective cross protective immunogenic antigen. It is widely considered as a diagnostic marker for leptospirosis. In this study, we evaluated the serodiagnostic potential of LipL21 protein of Leptospira interrogans serovar Pomona. We have successfully amplified, cloned, and expressed LipL21 in E. coli and evaluated its specificity by immunoblotting. Purified recombinant LipL21 (rLipL21) was inoculated into rabbits for the production of polyclonal antibody. Characterization of the purified IgG antibody against rLipL21 was performed by cross reactivity assay. Only sera from leptospirosis patients and rabbit hyperimmune sera recognized rLipL21 while the nonleptospirosis control sera showed no reaction in immunoblotting. We confirmed that anti-rLipL21-IgG antibody cross reacted with and detected only pathogenic leptospiral species and it did not react with nonpathogenic leptospires and other bacterial species. Results observed showed that anti-rLipL21-IgG antibody has high specificity and sensitivity to leptospires. The findings indicated that the antibody could be used in a diagnostic assay for detection of leptospires or their proteins in the early phase of infection.
  12. Fulltext Serological prevalence of leptospiral infection in wild rats at the National Service Training Centres in Kelantan and Terengganu
    Mohamed-Hassan SN, Bahaman AR, Mutalib AR, Khairani-Bejo S
    Trop Biomed, 2010 Apr;27(1):30-2.
    PMID: 20562810 MyJurnal
    One hundred and sixty eight rats were trapped from the National Service Training Centres (NSTC) in Kelantan and Terengganu from October 2008 to May 2009. Microscopic agglutination test (MAT) was performed to detect the presence of agglutinating antibodies to Leptospira among the rats caught. All the MAT positive rats were identified as Rattus tiomanicus. In Kelantan, 17.3 % (14/81) of the rats had leptospiral antibodies to serovars Icterohaemorrhagiae (12.3%), Canicola (2.5%), Ballum (1.2%), and Pyrogenes (1.2%). In Terengganu, 18.4% (16/87) of the rats had antibodies to serovars Icterohaemorrhagiae (15%), Canicola (1.1%), Pyrogenes (1.1%) and Hebdomadis (1.1%). This study indicated that Leptospira serovars were prevalent in the rat population in the study areas and could be a source of infection to humans. Therefore, control of the rat population in all NSTC is critical to prevent outbreaks of leptospirosis amongst the NSTC trainees.
  13. Prevalence of Escherichia Coli O157:H7 and Enterobacteriaceae on Hands of Workers in Halal Cattle Abattoirs in Peninsular Malaysia
    Shamsul BM, Adamu MT, Mohd Desa MN, Khairani-Bejo S
    Malays J Med Sci, 2016 Sep;23(5):65-71.
    PMID: 27904427
    BACKGROUND: Several occupational diseases of multiple origins are encountered among abattoir workers. Presence of indicator microorganisms (coliforms) on hands of workers can be used a gauge for hygienic practices.

    METHODS: A cross-sectional study was performed to assess the prevalence of E.coli and enterobacteriaceae among Halal abattoir workers in some government halal abattoirs of Malaysia. A total of one hundred and sixty-five hand swab samples were collected from workers of Halal abattoirs in Malaysia. The samples were subjected to microbiological analysis for characterisation and serotyping.

    RESULTS: The results have shown that no Escherichia coli O157:H7 was isolated on the hands of abattoir workers before and after work. However, a total prevalence of 9.7% was recorded for all samples during work. For non-O157:H7, total prevalence of 33.3% during work and 13% after work were obtained. High prevalence was recorded in sample taken during work from Tampin, Jasin and Kemaman (100% each) while low prevalence where observed in Shah Alam, Banting and Ipoh (20% each).

    CONCLUSIONS: Based on the findings the hygienic practices of hand washing among the workers in few locations was found to be low especially after work.

  14. Fulltext Identification of salmonella and other pathogenic bacteria in pet red-eared sliders, trachemys scripta elegans, from pet shops in the Klang valley
    Selvarajah, G.T., Khairani-Bejo, S.
    Jurnal Veterinar Malaysia, 2019;31(2):23-27.
    MyJurnal
    Bacteriological isolation and identification were performed on 60 cloacal swabs and 15 aquarium water samples of pet red-eared sliders (Trachemys scripta elegans) obtained from aquarium shops in the Klang Valley, Central Peninsula Malaysia. The most common bacteria isolated was Aeromonas spp., which was present in both cloacal swabs (70%) and aquarium water (86.7%). Klebsiella spp. (50%), Escherichia coli (33.3%), Yersinia spp. (16.7%) and Salmonella spp. (15%) obtained form cloacal swabs were identified as pathogenic to both humans and animals. Salmonella spp. were isolated from both cloacal swabs and aquarium water. The Salmonella serotypes identified were S. tennessee, S. typhimurium, S. brezany, S. pomona, S. corvallis and S. schwarzengrund. Bacterial infections in humans associated with handling exotic pets directly or indirectly in contact with aquarium water have been described regularly, hence the zoonotic significance of owning a turtle infected with Salmonella spp. or any pathogenic bacteria therefore cannot be ignored.
  15. Fulltext Risk Factors and Prediction of Leptospiral Seropositivity Among Dogs and Dog Handlers in Malaysia
    Goh SH, Ismail R, Lau SF, Megat Abdul Rani PA, Mohd Mohidin TB, Daud F, et al.
    Int J Environ Res Public Health, 2019 04 28;16(9).
    PMID: 31035316 DOI: 10.3390/ijerph16091499
    This study determined the potential risk factors that may contribute to seropositivity among dogs and dog handlers from working dog and dog shelter institutions. Data was collected from dogs (n = 266) and dog handlers (n = 161) using a standardised guided questionnaire. Serum obtained from the dogs and dog handlers was tested using the microscopic agglutination test (MAT). A logistic regression analysis was used to predict leptospiral seropositivity of dogs and dog handlers based on potential risk factors. A total of 22.2% of dogs and 21.7% of dog handlers were seropositive. The significant predictors for the dogs' seropositivity were presence of rats (OR = 4.61 (95% CI: 1.05, 20.33), p = 0.043) and shared common area (OR = 5.12 (95% CI: 1.94, 13.46), p = 0.001) within the organisation. Significant predictor for dog handler seropositivity was contact time with the dogs of more than six hours/day (OR = 3.28 (95% CI: 1.28, 8.40), p = 0.013) after controlling for the effect of other risk factors such as small mammal contact, rat infestation at home, flooding at housing area (within three months) and urban locality. The exposure to various disease sources identified poses risk to dogs and dog handlers. Risk could be reduced with adequate application of protection at work while handling dogs and thus limiting contact with these sources and reducing exposure to infection.
  16. Detection and distribution of anti-leptospiral antibody among dogs and their handlers
    Goh SH, Khor KH, Ismail R, Megat Abdul Rani PA, Mohd Mohidin TB, Bahaman AR, et al.
    Trop Biomed, 2020 Dec 01;37(4):1074-1082.
    PMID: 33612759 DOI: 10.47665/tb.37.4.1074
    The incidence of leptospirosis seems to be on the rise and could be an alarming indirect indication of a global re-emergence. It is a potential public health threat when dogs are speculated to be involved in the transmission of leptospirosis through possible subclinical harbouring of Leptospira spp. and subsequent shedding into the environment. This study aimed to detect anti-leptospiral antibodies among dogs and their handlers using the microscopic agglutination test (MAT). Blood samples from 266 apparently healthy dogs and 194 dog handlers were collected at four working dog organisations and four dog shelters. Serum samples were tested using MAT against 20 leptospiral serovars with a cut-off titre >=1:100 (dog) and >=1:50 (dog handlers). Seventy dogs (70/266; 26.3%) were seropositive mainly against serovars Icterohaemorrhagiae, Ballum, Bataviae and Javanica (titres ranged: 1:100-1:800). Sixty-seven dog handlers (67/194; 34.5%) were seropositive mainly against serovars Grippotyphosa, Icterohaemorrhagiae and Malaysia (titres ranged: 1:50-1:200). Dogs were seropositive due to exposure, vaccination or active infection. Seropositive dog handlers could indicate exposure or active infection. This shows the potential of dogs in maintaining and spreading the infection in Malaysia. Due to the occupational risk as a result of frequent contact with dogs and exposure to contaminated environments, dog handlers should be made aware of the presence of this zoonotic disease.
  17. Fulltext Detection of pathogenic Leptospira from selected environment in Kelantan and Terengganu, Malaysia
    Ridzlan FR, Bahaman AR, Khairani-Bejo S, Mutalib AR
    Trop Biomed, 2010 Dec;27(3):632-8.
    PMID: 21399605 MyJurnal
    Leptospirosis is recognized as one of the important zoonotic diseases in the world including Malaysia. A total of 145 soil and water samples were collected from selected National Service Training Centres (NSTC) in Kelantan and Terengganu. The samples were inoculated into modified semisolid Ellinghausen McCullough Johnson Harris (EMJH) medium, incubated at room temperature for 1 month and examined under the dark-field microscope. Positive growth of the leptospiral isolates were then confirmed with 8-Azaguanine Test, Polymerase Chain Reaction (PCR) assay and Microscopic Agglutination Test (MAT). Fifteen cultures (10.34%) exhibited positive growths which were seen under dark field microscope whilst only 20% (3/15) were confirmed as pathogenic species. based on 8-Azaguanine Test and PCR. Serological identification of the isolates with MAT showed that hebdomadis was the dominant serovar in Terengganu. Pathogenic leptospires can be detected in Malaysian environment and this has the potential to cause an outbreak. Therefore, precautionary steps against leptospirosis should be taken by camp authorities to ensure the safety of trainees.
  18. Fulltext Retrospective Study of Leptospirosis in Malaysia
    Garba B, Bahaman AR, Khairani-Bejo S, Zakaria Z, Mutalib AR
    Ecohealth, 2017 Jun;14(2):389-398.
    PMID: 28405850 DOI: 10.1007/s10393-017-1234-0
    Leptospirosis is a bacterial disease transmitted to humans and animals by direct or indirect contact with urine or body fluids from infected animals especially rodents. Infection can be associated with wide clinical spectrum varying from asymptomatic to severe multi-organ syndrome with life-threatening consequences. We conducted a review of published studies on incidences, case reports, sero-epidemiological surveys from year 2000 to 2015 using different electronic data bases. Our study revealed that majority of the studies were conducted in Peninsular Malaysia and predominantly among high-risk human groups. Most of the studies on domestic animals were conducted in the 1980s; hence, the current status of leptospirosis among domestic animal population remains largely unknown. There tend to be a sharp rise in incidence rate among human population in the year 2014 which was attributed to flooding and heavy rainfall experienced as well as recreational activities. Several gaps in epidemiological knowledge were also disclosed.
  19. Fulltext TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples
    Rahman MSA, Khor KH, Khairani-Bejo S, Lau SF, Mazlan M, Roslan MA, et al.
    J Vet Res, 2023 Jun;67(2):187-195.
    PMID: 38143826 DOI: 10.2478/jvetres-2023-0024
    INTRODUCTION: Canine leptospirosis has always been a differential diagnosis in dogs presenting with clinical signs and blood profiles associated with kidney and/or liver disease. The conventional polymerase chain reaction (PCR) provides diagnoses, but real-time PCR-based tests provide earlier confirmation and determine the severity of infection, especially in the acute stage, allowing early detection for immediate treatment decisions. To our knowledge, real-time PCR has not been routinely adopted for clinical investigation in Malaysia. This study evaluated TaqMan real-time PCR (qPCR) assays diagnosing leptospirosis and compared their applicability to clinical samples from dogs with kidney and/or liver disease against a conventional PCR reference.

    MATERIAL AND METHODS: The qPCR assays were validated using existing leptospiral isolates. Whole blood and urine samples were analysed using a conventional PCR, LipL32(1) and LipL32(2) qPCRs and a microscopic agglutination test. The sensitivity and specificity of the qPCRs were determined.

    RESULTS: The LipL32(1) qPCR assay had more diagnostic value than the LipL32(2) qPCR assay. Further evaluation of this assay revealed that it could detect as low as five DNA copies per reaction with high specificity for the tested leptospiral strains. No cross-amplification was observed with other organisms. Analysing the clinical samples, the LipL32(1) qPCR assay had 100.0% sensitivity and >75.0% specificity.

    CONCLUSION: The LipL32(1) qPCR assay is sensitive, specific and has the potential to be applied in future studies.

  20. Fulltext Serological diagnostic potential of recombinant outer membrane proteins (rOMPs) from Brucella melitensis in mouse model using indirect enzyme-linked immunosorbent assay
    Ahmed IM, Khairani-Bejo S, Hassan L, Bahaman AR, Omar AR
    BMC Vet Res, 2015;11:275.
    PMID: 26530141 DOI: 10.1186/s12917-015-0587-2
    Brucella melitensis is the most important pathogenic species of Brucella spp. which affects goats and sheep and causes caprine and ovine brucellosis, respectively. Serological tests for diagnosis of brucellosis such as Rose Bengal plate test (RBPT) and enzyme-linked immunosorbent assay (ELISA) usually utilize smooth lipopolysaccharides (S-LPS) as a diagnostic antigen which could give false positive serological reactions. Outer membrane proteins (OMP) of B. melitensis have been used as alternative diagnostic antigens rather than S-LPS for differential serological diagnosis of brucellosis, mainly in ELISA with single recombinant OMP (rOMP) as a diagnostic antigen. Nevertheless, the use of single format mainly showed lack of sensitivity against the desired rOMP. Therefore, this study aimed to determine whether a newly developed rOMPs indirect ELISA (rOMPs I-ELISA), based on combination of rOMP25, rOMP28 and rOMP31of B. melitensis, has a potential benefit for use in the serodiagnosis of brucellosis.
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