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Conformation sensitive gel electrophoresis for the detection of calreticulin mutations in BCR-ABL1-negative myeloproliferative neoplasms

Zakaria NA, Rosle NA, Siti Asmaa MJ, Aziee S, Haiyuni MY, Samat NA, et al.

Int J Lab Hematol, 2021 Dec;43(6):1451-1457.
PMID: 34125992 DOI: 10.1111/ijlh.13628

INTRODUCTION: Calreticulin (CALR) mutations in myeloproliferative neoplasms (MPN) have been reported to be key markers in the molecular diagnosis, particularly in patients lacking JAK2 V617F mutation. In most current reports, CALR mutations were analysed by either allele-specific PCR (AS-PCR), or the more expensive quantitative real-time PCR, pyrosequencing and next-generation sequencing. Hence, we report the use of an alternative method, the conformation sensitive gel electrophoresis (CSGE) for the detection of CALR mutations in BCR-ABL1-negative MPN patients.
METHODS: Forty BCR-ABL1-negative MPN patients' DNA: 19 polycythemia vera (PV), 7 essential thrombocytosis (ET) and 14 primary myelofibrosis (PMF), were screened for CALR mutations by CSGE. PCR primers were designed to amplify sequences spanning between exons 8 and 9 to target the mutation hotspots in CALR. Amplicons displaying abnormal CSGE profiles by electrophoresis were directly sequenced, and results were analysed by BioEdit Sequence Alignment Editor v7.2.6. CSGE results were compared with AS-PCR and confirmed by Sanger sequencing.
RESULTS: CSGE identified 4 types of mutations; 2 PMF patients with either CALR type 1 (c.1099_1150del52) or type 2 (c.1155_1156insTTGTC), 1 ET patient with nucleotide deletion (c.1121delA) and insertion (c.1190insA) and 1 PV patient with p.K368del (c.1102_1104delAAG) and insertion (c.1135insA) inframe mutations. Three patients have an altered KDEL motif at the C-terminal of CALR protein. In comparison, AS-PCR only able to detect two PMF patients with mutations, either type 1 and type 2.
CONCLUSION: CSGE is inexpensive, sensitive and reliable alternative method for the detection of CALR mutations in BCR-ABL1-negative MPN patients.
Fulltext Effect of incorporating sodium molybdate in the form of salt lick or in mixed ration on growth and performance of sheep fed palm kernel cake

Yusoff SM, Ibrahim J, Salleh A

Malays J Nutr, 1995 Sep;1(2):171-8.
PMID: 22692061

The objective of the present experiments are to determine if sheep could safely consume high amount of PKC in their diets and if sheep's consumption of PKC require a chelating agent to tie up the high copper level in PKC to protect it against toxicity. Two experiments were carried out. Experiment 1, with four treatment groups of animals, using from 30 to 100% PKC, mixed with other feed ingredients except minerals. Mineral mixtures were separately mixed with Sodium Molybdate, acting as the chelating agent, and the mineral was offered in separate feed troughs ad.libitum, in the pens for each animal group. Although all groups gave high ADG, the animals in the 100% group had high copper in their blood, which were above4 the normal physiological level at the end of the experimental duration of three months. Three animals from this group died and their liver copper contents were very high. The groups fed up to 72% PKC in their ration did not show any significant elevation of copper or toxicity. Sheep fed similar proportions of ingredients in experiment 2, but with Molybdate incorporated together into all the four similar rations as in experiment 1, did not show any signs of toxicity or elevated blood copper. The animals in all groups produced high ADG. The experiment proved that sheep can take up to 100% PKC in their diet, but a chelating agent must be incorporated into the feed to ensure its sufficient uptake to protect it against toxicity. Giving Molybdate separately from the feed would not ensure sufficient intake of the chelating agent voluntarily. This would result in copper toxicity in the animals.
Fulltext Initial evaluation of the training programme for health care professionals on the use of Malaysian clinical practice guidelines for management of dementia

Yusoff S, Koh CT, Mohd Aminuddin MY, Krishnasamy M, Suhaila MZ

East Asian Arch Psychiatry, 2013 Sep;23(3):91-101.
PMID: 24088402

The Malaysian Clinical Practice Guidelines (CPG) for Management of Dementia (second edition) was launched in April 2010 by the Ministry of Health Malaysia. A training programme for the management of dementia, involving all categories of staff working at primary and secondary centres, was implemented to ensure that care delivery for people with dementia was in accordance with the guidelines. The study aimed to look into improving knowledge and understanding of dementia following training, and to evaluate the effectiveness of the training programme using a clinical audit indicator recommended in the guidelines.
Frequencies of A(TA)7TAA, G71R, and G493R mutations of the UGT1A1 gene in the Malaysian population

Yusoff S, Van Rostenberghe H, Yusoff NM, Talib NA, Ramli N, Ismail NZ, et al.

Biol. Neonate, 2006;89(3):171-6.
PMID: 16210851

Gilbert syndrome is caused by defects in the uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) gene. These mutations differ among different populations and many of them have been found to be genetic risk factors for the development of neonatal jaundice.
Increased Expression of Phosphatidylinositol 3-Kinase p110α and Gene Amplification of PIK3CA in Nasopharyngeal Carcinoma

Yip WK, He PY, Abdullah MA, Yusoff S, Seow HF

Pathol Oncol Res, 2016 Apr;22(2):413-9.
PMID: 26581613 DOI: 10.1007/s12253-015-0007-8

Molecular alterations in PIK3CA oncogene that encodes the p110α catalytic subunit of phosphatidylinositol 3-kinase (PI3K p110α) are commonly found in human cancers. In this study, we examined the expression of PI3K p110α and PIK3CA gene amplification in 74 nasopharyngeal carcinoma (NPC) cases. Immunohistochemical staining demonstrated overexpression of PI3K p110α protein in 44.6% (33/74) of NPCs and 4.8% (2/42) of the adjacent normal nasopharyngeal mucosa. Copy number of PIK3CA gene was successfully analyzed in 51 of the total NPC cases and 19 non-malignant nasopharynx tissues by quantitative real-time PCR. Using mean + 2(standard deviation) of copy numbers in the non-malignant nasopharynx tissues as a cutoff value, PIK3CA copy number gain was found in 10 of 51 (19.6%) NPC cases. High PI3K p110α expression level was correlated with increased PIK3CA copy number (Spearman's rho =0.324, P = 0.02). PI3K p110α expression and PIK3CA copy number did not associate with Akt phosphorylation, and patient and tumor variables. This study suggests that PI3K p110α overexpression, which is attributed, at least in part, to PIK3CA gene amplification, may contribute to NPC pathogenesis. However, these molecular aberrations may not be responsible for activation of Akt signaling in NPC.
Overexpression of phospho-Akt correlates with phosphorylation of EGF receptor, FKHR and BAD in nasopharyngeal carcinoma

Yip WK, Leong VC, Abdullah MA, Yusoff S, Seow HF

Oncol Rep, 2008 Feb;19(2):319-28.
PMID: 18202777

The Akt pathway is one of the most common molecular alterations in various human malignancies. However, its involvement in nasopharyngeal carcinoma (NPC) tumorigenesis has not been well established. In this study, the status of Akt activation and expression of its upstream and downstream molecules was investigated in 64 NPC and 38 non-malignant nasopharyngeal tissues by immunohistochemistry. The hotspot mutations of PIK3CA, encoding the p110alpha catalytic subunit of phosphatidylinositol 3-kinase (PI3K), were also determined in 25 of these NPC tissues. No hotspot mutations were found in any of the samples tested. Akt was activated in 27 (42.2%) and 23 (35.9%) NPCs, as indicated by p-Akt (Thr308) and p-Akt (Ser473) immunoreactivity, respectively. PTEN loss did not correlate statistically with activated Akt. However, a positive correlation was observed between activated Akt and phospho-epidermal growth factor receptor (p-EGFR), suggesting that the EGFR signaling might be one of the upstream regulators of the Akt pathway. The phosphorylation of forkhead (FKHR) and Bcl-2 associated death domain (BAD), but not mammalian target of rapamycin and glycogen synthase kinase-3beta, was significantly correlated with Akt activation. This implies that Akt promotes cell proliferation (as estimated by Ki-67) and survival, at least, through the inactivation of FKHR and BAD in NPC. Our data revealed that the EGFR/PI3K/Akt signaling pathway is important in NPC pathogenesis and that PIK3CA hotspot mutations are rare in NPC.
Fulltext Increase in tumour-infiltrating lymphocytes with regulatory T cell immunophenotypes and reduced zeta-chain expression in nasopharyngeal carcinoma patients

Yip WK, Abdullah MA, Yusoff SM, Seow HF

Clin Exp Immunol, 2009 Mar;155(3):412-22.
PMID: 19220831 DOI: 10.1111/j.1365-2249.2008.03793.x

The pathological significance of the mechanisms of tumour immune-evasion and/or immunosuppression, such as loss of T cell signalling and increase in regulatory T cells (T(regs)), has not been well established in the nasopharyngeal carcinoma (NPC) microenvironment. To evaluate the T(reg) immunophenotypes in tumour-infiltrating lymphocytes (TILs), we performed a double-enzymatic immunostaining for detection of forkhead box P3 (FoxP3) and other markers including CD4, CD8, and CD25 on 64 NPC and 36 non-malignant nasopharyngeal (NP) paraffin-embedded tissues. Expression of CD3 zeta and CD3 epsilon was also determined. The prevalence of CD4(+)FoxP3(+) cells in CD4(+) T cells and the ratio of FoxP3(+)/CD8(+) were increased significantly in NPC compared with those in NP tissues (P < 0.001 and P = 0.025 respectively). Moreover, the ratio of FoxP3(+)/CD25(+)FoxP3(-) in NPC was significantly lower than that in NP tissues (P = 0.005), suggesting an imbalance favouring activated phenotype of T cells in NPC. A significant negative correlation between the abundance of FoxP3(+) and CD25(+)FoxP3(-) cells (P < 0.001) was also identified. When histological types of NPC were considered, a lower ratio of FoxP3(+)/CD25(+)FoxP3(-) was found in non-keratinizing and undifferentiated carcinomas. Increased CD4(+)FoxP3(+)/CD4(+) proportion and FoxP3(+)/CD8(+) ratio were associated with keratinizing squamous cell carcinoma. A reduced expression of CD3 zeta in TILs was found in 20.6% of the NPC tissues but none of the NP tissues. These data provide evidence for the imbalances of T(reg) and effector T cell phenotypes and down-regulation of signal-transducing molecules in TILs, supporting their role in suppression of immune response and immune evasion of NPC.
Establishment of suitable separation distance by using different methods for malodor mitigation from palm oil mill

Yaacof N, Qamaruz Zaman N, Yusup Y, Yusoff S

Environ Sci Pollut Res Int, 2019 Aug;26(23):24286-24299.
PMID: 31214886 DOI: 10.1007/s11356-019-05517-z

Malaysia is the second-largest producer and exporter of palm oil amounting to 39% of world palm oil production and 44% of world exports (MPOB, 2014). An enormous amount of palm oil mill effluent is released during palm oil milling, and the effluent causes a major odor problem. Many methods, such as biofiltering, can be adopted to manage the malodor. However, these methods are expensive and require high maintenance. The separation distance method can be used as an alternative due to its low cost and effectiveness. This research was conducted to verify the performance of three different methods, namely, in-field monitoring by using an olfactometer, CALPUFF model, and Gaussian plume model. Given that no research has compared the three methods, this study examined the effectiveness of the methods and determined which among them is suitable for use in Malaysia. The appropriate separation distances were 1.3 km for in-field monitoring, 1.2 km for the CALPUFF model, and 0.5 for the Gaussian plume model. These different values of separation distance were due to the various approaches involved in each method. This research determined an appropriate means to establish a proper separation distance for reducing odor nuisance in areas around palm oil mills.
Fulltext Clinical predictors of acute kidney injury in children with acute post-streptococcal glomerulonephritis: a tertiary centre experience

Wan Yusof WA, Yaacob NM, Nasir A, Yusoff S, Ilias MI

Singapore Med J, 2023 Jan 06.
PMID: 36695277 DOI: 10.4103/singaporemedj.SMJ-2021-092
Fulltext Potential use of cord blood for Hb E hemoglobinopathy screening programme using capillary electrophoresis

Wan Mohd Saman WA, Hassan R, Mohd Yusoff S, Che Yaakob CA, Abdullah NA, Ghazali S, et al.

Malays J Pathol, 2016 Dec;38(3):235-239.
PMID: 28028293 MyJurnal

BACKGROUND: Thalassemia and hemoglobinopathies are inherited red blood cell disorders found worldwide. Hemoglobin (Hb) E disorder is one of the hemoglobinopathies known to have the high prevalence in South East Asia. Most of transfusion-dependent thalassemias were genotypically compound heterozygous Hb E/ β-thalassemia. In Malaysia, the national screening program for thalassemia was implemented for early pregnancy or secondary school girls; however many participants do not turn-up and missed the screening test. Screening for thalassemia using samples from cord blood is an alternative choice as it is a readily available source of blood and hence early detection of the disease. The purpose of this study was to determine the potential use of cord blood for the screening of HbE hemoglobinopathy by using capillary electrophoresis (CE).
METHODS: Cord blood samples were collected from 300 newborns of healthy mothers. Hematological parameters were determined and hemoglobin quantitation for all cord blood samples were performed using capillary electrophoresis system (CES) and high performance liquid chromatography (HPLC).
RESULTS: Majority of cord blood samples (63%) revealed Hb AF followed by Hb AFA2 (20%). Hb AFE was detected in 10.7% with the mean value of Hb E ranging from 2.3%-11.1%.
CONCLUSION: Hemoglobin E was detected in cord blood using capillary electrophoresis system. It can be recommended in areas where Hb E/β is prevalent. Implementation of a screening strategy using CE on cord blood sampling will identify the disease early. With regular follow-up on these patients, the status of their disease can be determined earlier and appropriate management implemented.
Effects of resveratrol on shedding and pathological dynamics in experimental B. melitensis infection in dexamethasone-treated nonpregnant Boer goats

Tanko PN, Mohd Yusoff S, Emikpe BO, Bejo SK, Salisi S

J Immunoassay Immunochem, 2019;40(4):419-438.
PMID: 31154897 DOI: 10.1080/15321819.2019.1620766

Brucellosis constitutes an infectious re-emerging zoonosis. Spread of diseases could be exacerbated by stress-induced immunosuppression. This study evaluated relationship between Brucella melitensis infection, shedding dynamics, dexamethasone-induced stress, pathological alterations and resveratrol ameliorative effects in goats. Twelve nonpregnant goats were divided into four groups A, B, C, and D of three animals each. Groups A and B were administered 107 CFU/mL of B. melitensis ocularly, 21 days prior to 7 days consecutive administration of dexamethasone (2 mg/kg). Group A was further administered resveratrol (5 mg/kg) intravenously for 5 consecutive days from day 31 post B. melitensis inoculation. Group C was administered similar dose of B. melitensis while group D was inoculated normal saline. Blood, nasal, ocular, and vaginal swabs were collected at intervals for analysis. The does were sacrificed at day 42 post inoculation (pi). Tissues were collected for tissue bacterial load determination, histopathology, and immunohistochemistry. Dexamethasone administration from day 21 pi increased the frequency in the shedding dynamics, tissue bacterial load, pathological alterations (frequency of microgranuloma and intensity of immunostaining) in group B while 5 days treatment with resveratrol following dexamethasone administration significantly reduced tissue bacterial load, decline in shedding dynamics, and ameliorate damage by dexamethasone administration/B. melitensis infection.
Dexamethasone-induced stress exacerbates shedding, tissue antigen distribution, and pathology of caprine Brucellosis

Tanko P, Mohd Yusoff S, Emikpe BO, Onilude OM, Abdullateef A

J Immunoassay Immunochem, 2021 May 04;42(3):265-284.
PMID: 33577382 DOI: 10.1080/15321819.2020.1862862

This study investigated dexamethasone-treatment, shedding routes, tissue antigen distribution, and pathology of caprine Brucellosis. Eighteen non-pregnant goats were randomly grouped into A, B, and C. Group A was administered dexamethasone for 7 days at 2 mg/kg before inoculating 0.5 mL B. melitensis at 107 CFU ocularly while group B was inoculated 0.5 mL B. melitensis only, and C as control negative. Blood samples, ocular, nasal, and vaginal swabs were obtained for evaluation. Three goats were sacrificed from each group at days 21 and 42 post-inoculation (pi) and selected tissues collected for PCR, histopathology, and immunohistochemistry. Brucella melitensis was detected in the ocular swabs of group A significantly higher than group B. Shedding was prolonged in group A compared to B. The overall shedding was 22.2% in group A and 9.4% in group B. The uterus of both groups A and B revealed mild inflammation and microgranuloma, extensive necrotic lesions in lymph nodes. Liver showed multifocal necrosis predominantly in group A. Lesion scoring showed significantly higher scores in A compared to B. Strong immunostaining was observed in the liver, lungs, and spleen, predominantly at day 21 pi. This study demonstrated dexamethasone prolonged shedding, tissue antigen distribution, and pathology in dexamethasone-treated goats.
A Rare Case of an Infant with Left Hemiparesis: A Case Report of Bilateral Open-lip Schizencephaly

Siti BC, Zulkifli MM, Mohd Yusoff SS, Muhamad R, Ahmad TM

Malays Fam Physician, 2020;15(3):90-94.
PMID: 33329869

Schizencephaly is a very rare congenital birth defect. It is characterized by a cortical brain malformation that manifests as a grey-matter-lined cleft extending from the ependyma to the pia mater. It is a rare condition, and few cases have been reported in the literature. The exact cause is unknown. Herein, we report a case of an infant presenting with left side hemiparesis. The CT scan of her brain revealed right fronto-temporal and left parieto-temporal open-lip schizencephaly; thus, urgent referral to a pediatric neurologist was made for early intervention.
Cyclospora sp causing diarrhea in man

Sinniah B, Rajeswari B, Johari S, Ramakrishnan K, Yusoff SW, Rohela M

Southeast Asian J. Trop. Med. Public Health, 1994 Mar;25(1):221-3.
PMID: 7825021
Screening of the LIX1 gene in Japanese and Malaysian patients with SMA and/or SMA-like disorder

Sasongko TH, Gunadi, Yusoff S, Atif AB, Fatemeh H, Rani A, et al.

Brain Dev, 2010 May;32(5):385-9.
PMID: 19664890 DOI: 10.1016/j.braindev.2009.06.008

The majority of spinal muscular atrophy (SMA) patients showed homozygous deletion or other mutations of SMN1. However, the genetic etiology of a significant number of SMA patients has not been clarified. Recently, mutation in the gene underlying cat SMA, limb expression 1 (LIX1), has been reported. Similarity in clinical and pathological features of cat and human SMA may give an insight into possible similarity of the genetic etiology.
High allele frequency of CYP2C9*3 (rs1057910) in a Negrito's subtribe population in Malaysia; Aboriginal people of Jahai

Rosdi RA, Mohd Yusoff N, Ismail R, Soo Choon T, Saleem M, Musa N, et al.

Ann Hum Biol, 2015 Sep 24.
PMID: 26402341

CYP2C9 gene polymorphisms modulate inter-individual variations in the human body's responses to various endogenous and exogenous drug substrates. To date, little is known about the CYP2C9 gene polymorphisms among the aboriginal populations of the world, including those in Malaysia.
Fulltext A minireview of CYP2C9 and CYP2C19 single nucleotide polymorphisms (SNPs) among Malaysian populations

Rosdi R.A., Yusoff S., Mohd Yusoff N., Ismail R., Tan, C.S., Musa N.

Journal of Biomedical and Clinical Sciences, 2017;2(1):47-52.
MyJurnal

It has been recognized extensively that studies of pharmacogenetics provide massive examples of causal relationship between genotypes and drug effectiveness to account for interindividual phenotypic variations in drug therapy. In most cases, cytochrome P450 (CYP) polymorphisms are one of the major variables that affecting those drug plasma concentration, drug detoxification and drug activation in humans. Thus, understanding of CYP polymorphisms can be crucially valuable in order to allow early and more accurate drug dosage prediction and improve the drug response accordingly. Despite the high level of homologous amino acid sequences, CYP2C9 and CYP2C19 genes are among the most important CYP genes which metabolize a wide range of clinically therapeutic drugs. Several critical reviews have been published relating to the aforementioned genes. However, this minireview aims to systematically merge reported studies on the SNPs frequencies of both genes concentrating only on Malaysian population. It is hoped that, the minireview can be an opener for new opportunities to reevaluate the evidence on the prevalence of CYP2C genes as a potential genetic factor influencing a particular drug efficacy and safety among Malaysian. Such evaluation can be developed to the next level of early prediction of better and specific drug treatment, thereby improving the drug response while helping the government in minimising the drug expenditures.
Optimal implementation of low impact development for urban stormwater quantity and quality control using multi-objective optimization

Rezaei AR, Ismail Z, Niksokhan MH, Dayarian MA, Ramli AH, Yusoff S

Environ Monit Assess, 2021 Mar 31;193(4):241.
PMID: 33791871 DOI: 10.1007/s10661-021-09010-4

Stormwater runoff is a major concern in urban areas which is mostly the result of vast urbanization. To reduce urban stormwater runoff and improve water quality, low impact development (LID) is used in urban areas. Therefore, it is vital to find the optimal combination of LID controls to achieve maximum reduction in both stormwater runoff and pollutants with optimal cost. In this study, a simulation-optimization model was developed by linking the EPA Storm Water Management Model (SWMM) to the Multi-Objective Particle Swarm Optimization (MOPSO) using MATLAB. The coupled model could carry out multi-objective optimization (MOO) and find potential solutions to the optimization objectives using the SWMM simulation model outputs. The SWMM model was developed using data from the BUNUS catchment in Kuala Lumpur, Malaysia. The total suspended solids (TSS) and total nitrogen (TN) were selected as pollutants to be used in the simulation model. Vegetated swale and rain garden were selected as LID controls for the study area. The LID controls were assigned to the model using the catchment characteristics. The target objectives were to minimize peak stormwater runoff, TSS, and TN with the minimum number of LID controls applications. The LID combination scenarios were also tested in SWMM to identify the best LID types and combination to achieve maximum reduction in both peak runoff and pollutants. This study found that the peak runoff, TSS, and TN were reduced by 13%, 38%, and 24%, respectively. The optimal number of LID controls that could be used at the BUNUS catchment area was also found to be 25.
Coastal meteorology on the dispersion of air particles at the Bachok GAW Station

Rahim HA, Khan MF, Ibrahim ZF, Shoaib A, Suradi H, Mohyeddin N, et al.

Sci Total Environ, 2021 Aug 15;782:146783.
PMID: 33838363 DOI: 10.1016/j.scitotenv.2021.146783

Meteorology over coastal region is a driving factor to the concentration of air particles and reactive gases. This study aims to conduct a research to determine the level of year-round air particles and the interaction of the meteorological driving factors with the particle number and mass in 2018, which is moderately influenced by Southeast Asian haze. We obtained the measurement data for particle number count (PNC), mass, reactive gases, and meteorological factors from a Global Atmospheric Watch (GAW) station located at Bachok Marine Research Center, Bachok, Kelantan, Malaysia. For various timeseries and correlation analyses, a 60-second resolution of the data has been averaged hourly and daily and visualized further. Our results showed the slight difference in particle behavior that is either measured by unit mass or number count at the study area. Diurnal variations showed that particles were generally high during morning and night periods. Spike was observed in August for PM2.5/PNC2.5 and PM10/PNC10 and in November for PMCoarse/PNCCoarse. From a polar plot, the particles came from two distinct sources (e.g., seaside and roadside) at the local scale. Regional wind vector shows two distinct wind-blown directions from northeast and southwest. The air mases were transported from northeast (e.g., Philippines, mainland China, and Taiwan) or southwest (e.g., Sumatra) region. Correlation analysis shows that relative humidity, wind direction, and pressure influence the increase in particles, whereas negative correlation with temperature is observed, and wind speed may have a potential role on the decline of particle concentration. The particles at the study area was highly influenced by the changes in regional wind direction and speed.
Development and application of dot-enzyme-linked immunosorbent (dot-ELISA) assay for detection of Brucella melitensis and evaluation of the shedding pattern in infected goats

Onilude OM, Mohd Yusoff S, Emikpe BO, Tanko P, Shahrom SM, Effendy M

J Immunoassay Immunochem, 2017;38(1):82-99.
PMID: 27494045 DOI: 10.1080/15321819.2016.1220395

Early and accurate diagnosis of Brucella melitensis is essential for the treatment and control of brucellosis both in animals and humans. The thrust for the development of a rapid diagnostic technique to overcome the limitations of conventional microbiological and serological tests brought about this investigation on the development and application of dot-ELISA for antigen and antibody detection in infected goats. Fifteen apparently healthy Boer aged 2-3 years which tested negative for brucellosis using PCR and ELISA, were grouped into A (10 goats infected intraocularly with 10(7) CFU of B. melitensis) and B (5 goats) as control. Discharges (ocular, nasal, and vaginal) and blood were collected at days 3, 7, 10, 14, weekly until 42 post-infection (pi) for dot-ELISA, PCR, and RBPT. Dot-ELISA detected B. melitensis antigen and antibody in group A at day 3 and 7 pi, respectively with adequate sensitivity and specificity relative to PCR and RBPT. The bacteria shedding detected from discharges at day 3 pi in the nasal and ocular route with dot-ELISA. Group B were consistently negative. Values such as speed, simplicity, field adaptability, high sensitivity, and specificity make dot-ELISA a rapid and adequate technique for diagnosis of brucellosis in B. melitensis infected goats within few hours.

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