Rapid Agglutination Research Articles

Blood transfusions in dogs and cats: blood typing and cross matching

The blood transfusion (administration of blood) is a resource of veterinary medicine with several indications. Point-of-care blood-typing methods, including both typing cards and rapid gel agglutination, are readily available. Following blood typing, cross matching is performed on one or more donor units of appropriate blood type. Cross matching reduces the risk of transfusion reactions but does not completely eliminate the risk of other types of transfusion reactions in veterinary patients. All transfusion reactions should be appropriately documented and investigated. It is, therefore, important to have in-depth technical knowledge about concepts and procedures involved in clinical blood transfusion, so that the entire procedure can be performed in the most appropriate way and with a high success rate.

Incidence and risk factors of Mycoplasma synoviae infection in broiler breeder farms of Iran

Mycoplasma synoviae (MS) is an important pathogen of poultry worldwide, causing respiratory tract infection and infectious synovitis in chickens and turkeys. The aim of this study was to determine the incidence and risk factors (age, size of flock, locale, sex and strain) of Mycoplasma synoviae in broiler breeder farms in Iran. The study was based on Rapid Serum Plate Agglutination (SPA) and enzyme linked immunosorbent assay (ELISA) tests. The prevalence was highest (44%) in winter and lowest (34%) in summer. Ross, Cobb, Arian, Hubbard and Arbor Acres strains had 40%, 44%, 32%, 27% and 45% infection, respectively. The prevalence was recorded highest in above 60 weeks of age (47.8%), but at 10-20 weeks it was lowest (14.2%). No significant difference was seen in flocks up to 30,000 population (41.9%), 30,000-40,000 (52%) and upper 40,000 (57%). The prevalence of Mycoplasmosis in foothills was significantly (p<0.05) higher (41%) than coastal area (34.5%). The results showed that occurrence of MS have a significant relationship with the age and zone of sampling.

Sero-prevalence of hepatitis B virus infection and its risk factors among pregnant women attending antenatal clinic at Aminu Kano teaching hospital, Kano, Nigeria

Background: Pregnant women infected with hepatitis B virus (HBV) can transmit the infection to their fetuses and newborns. Neonates who contract the HBV have about 90% risk of developing chronic HBsAg carriage (HBsAg: hepatitis B surface antigen) and chronic liver disease. Neonatal immunization interrupts this vertical and perinatal transmission. Objectives: To determine the seroprevalence of HBsAg among pregnant women attending the antenatal clinic at Aminu Kano Teaching Hospital (AKTH) and to identify potential risk factors associated with HBV infection. Materials and Methods: A case control study was conducted involving a total of 303 pregnant women attending the antenatal clinic at AKTH and 303 nonpregnant women of childbearing age. Blood sample was collected from each woman and the serum tested for the presence of HBsAg using latex rapid agglutination slide test kit (Cal-Tech Diagnostic Inc., USA) in the laboratory of the hospital. Reactive samples were stored at -20oC and further confirmed for HBsAg using enzyme-linked immunosorbent assay (ELISA) kits (Bio-Rad, France). HBsAg-positive samples were tested for hepatitis B e antigen (HBeAg) using ELISA kits (Orgenics, Israel). A pretested, structured questionnaire was used for the collection of sociodemographic data and possible risk factors. Results: The prevalence of HBsAg among pregnant women and nonpregnant women were 7.9 and 7.6%, respectively. There was no statistically significant difference in the prevalence of HBsAg in pregnant and nonpregnant women. The presence of HBeAg was statistically significant among both pregnant and nonpregnant women who tested positive for HBsAg. The risk factors associated with HBV infection were blood transfusion, ear piercing, history of an affected sibling with HBV infection, tattooing, and abortion among pregnant women. Conclusion: The prevalence of HBsAg in this study was not statistically different in pregnant and nonpregnant women. There was a high level of HBeAg infection among pregnant women who tested positive for HBsAg. History of an affected sibling with HBV infection, tattoo, and abortion were significant risk factors for HBV infection. Keywords: Africa, hepatitis B, pregnancy, risk factors

Isolation, characterization and antibiotic susceptibility profile of Staphylococcus aureus from raw milk samples in Nagpur district, India

The study was carried out from July 2010 to June 2011. The aim of the present investigation was to isolate Staphylococcus aureus from raw milk obtained from cow from different parts of Nagpur district, India and to determine their antibiotic susceptibility profile. General biochemical tests along with the MASTASTAPHTM rapid agglutination tests were employed for bacterial identification. All the well characterized isolates were further confirmed by amplifying coa gene by PCR. A total 95 S. aureus isolates were obtained during this study from 50 raw milk samples. When all the well characterized isolates were subjected to antibiotic susceptibility testing, erythromycin and chloramphenicol were the most effective antibiotics. A large proportion of isolates were found to be resistant to Co-trimoxazole (89%) followed by Linomycin (81%) and Penicillin (35%) It was concluded from the current study that the hygiene of the milk was poor and resistant strains contaminated the milk probably during the process of transportation.

Rapid serum agglutination and agar gel immunodiffusion tests associated to clinical signs in rams experimentally infected with Brucella ovis

The purpose of this study was to evaluate the agar gel immunodiffusion and the rapid serum agglutination tests associated to clinical signs in rams experimentally infected with Brucella ovis. The serological profile during the 12 months of infection showed a large fluctuation of antibodies that favors the failure in the diagnostic. The evaluation of tests after the experimental infection allowed to suggest that none of the tests were able to detect the infection throughout the period of study. The study reinforces the importance of considering the clinical signs to support the diagnosis of Brucella ovis infection in rams.

Identification of Potential Serodiagnostic and Subunit Vaccine Antigens by Antibody Profiling of Toxoplasmosis Cases in Turkey

Toxoplasmosis, caused by infection of the protozoan parasite Toxoplasma gondii, is associated with mild disease in healthy individuals, whereas individuals with depressed immunity may develop encephalitis, neurologic disorders, and other organ diseases. Women who develop acute toxoplasmosis during pregnancy are at risk of transmitting the infection to the fetus, which may lead to fetal damage. A diagnosis is usually confirmed by measuring IgG, or IgM where it is important to determine the onset of infection. A negative IgM result essentially excludes acute infection, whereas a positive IgM test is largely uninterpretable because IgM can persist for up to 18 months after infection. To identify antigens for improved diagnosis of acute infection, we probed protein microarrays displaying the polypeptide products of 1357 Toxoplasma exons with well-characterized sera from Turkey. The sera were classified according to conventional assays into (1) seronegative individuals with no history of T. gondii infection; (2) acute infections defined by clinical symptoms, high IgM titers, and low avidity IgG; (3) chronic/convalescent cases with high avidity IgG but persisting IgM; (iv) true chronic infections, defined by high avidity IgG and no IgM. We have identified 38 IgG target antigens and 108 IgM target antigens that can discriminate infected patients from healthy controls, one or more of which could form the basis of a 'tier-1' test to determine current or previous exposure. Of these, three IgG antigens and five IgM antigens have the potential to discriminate chronic/IgM persisting or true chronics from recent acutely infected patients (a 'tier-2' test). Our analysis of the antigens revealed several enriched features relative to the whole proteome, which include transmembrane domains, signal peptides, or predicted localization at the outer membrane. This is the first protein microarray survey of the antibody response to T. gondii, and will help in the development of improved serodiagnostics and vaccines.

Isolation ofMycoplasma meleagridisfrom Chickens

Mycoplasma meleagridis (MM) is a major cause of disease and economic loss in turkeys. Formerly it was thought that this species was very host specific and only restricted to turkey. In this study, we report on the recovery of MM from breeding flocks of chickens located near a turkey breeding unit. Ten MM field strains were isolated (by culture on Frey broth medium) from tracheal swabs of chickens displaying clinical signs of mycoplasmosis-essentially respiratory symptoms and poor performance. Assignment of the isolated field strains to MM was confirmed by a growth inhibition assay using MM-specific polyclonal antiserum and by PCR amplification targeting the 16S rRNA sequence as well as the Mm14 sequence, a MM-species-specific DNA fragment previously identified and characterized in our laboratory. The nucleotide sequence of Mm14 proved to be highly conserved among the 10 MM field strains, indicating a common source of infection. However, on the basis of slight differences in sodium dodecyl sulfate-polyacrylamide gel electrophoresis whole-cell proteins and western blot profiles, two groups of the isolated MM field strains could be distinguished. Evidence of MM infection of chickens was further provided by serology, since 13.77% (35/254) of sera proved positive to MM by either rapid serum agglutination or recombinant antigen-based enzyme-linked immunosorbent assay. In addition, sera of all chickens from which MM was isolated were positive for antibodies to MM. Collectively, the data unambiguously show that MM could infect chickens; thus, MM warrants further exploration to determine its pathogenicity in this unusual host.

Development and immunogenicity of recombinant GapA+Mycoplasma gallisepticum vaccine strain ts-11 expressing infectious bronchitis virus-S1 glycoprotein and chicken interleukin-6

Mycoplasma gallisepticum (MG) is a major pathogen of poultry that causes chronic respiratory disease in chickens and infectious sinusitis in turkeys. A live attenuated vaccine, ts-11, has been used for the control of MG in several countries. The efficacy of this vaccine is highly dose dependent and the flock antibody response is weak. To improve the functionality of the vaccine and investigate its potential as a delivery vector for foreign antigens and immunomodulatory proteins, we developed a derivative of ts-11 expressing infectious bronchitis virus-S1 glycoprotein (IBV-S1) and releasing chicken interleukin-6 into the extracellular milieu (MG ts-11 C3 (+CS)) using a transposon-based delivery vector. Following administration of MG ts-11 C3 (+CS) to chickens by eye-drop, an antibody response to MG and IBV-S1, as determined by the rapid serum agglutination test (RSA) and Western blotting, respectively, could be detected. Birds inoculated with the recombinant vaccine had significantly enhanced weight gain and were partially protected against damage by pathogenic IBV. These results indicate that the ChIL-6 released by MG ts-11 C3 (+CS) may have had a non-specific effect on growth rate. They also suggest that ts-11 is a promising vaccine vector, capable of delivering heterologous protective antigens, and may also provide non-specific benefits when engineered to express immunomodulatory proteins. With some improvements in the expression system, it could be used to induce a targeted immune response against specific mucosal pathogens, and co-expression of several antigens would allow development of a novel multivalent vaccine.

Submicron Polymer Particles Containing Fluorescent Semiconductor Nanocrystals Cdse/Zns for Bioassays

This study aimed to design a panel of uniform particulate biochemical reagents and to test them in specific bioassays. These reagents are polymer particles of different sizes doped with semiconductor nanocrystals and conjugated with either full-size antibodies or recombinant mini-antibodies (4D5 scFv fragment) designed by genetic engineering approaches. A panel of highly fluorescent polymer particles (150-800 nm) were formed by embedding CdSe/ZnS nanocrystals (quantum dots) into preformed polyacrolein and poly(acrolein-co-styrene) particles. Morphology, content and fluorescence characteristics of the prepared materials were studied by laser correlation spectroscopy, spectrophotometry, optical and fluorescent microscopy and fluorimetry. The obtained fluorescent particles sensitized by anti-Yersinia pestis antibodies were used for rapid agglutination glass test suitable for screening analysis of Y. pestis antigen and for microtiter particle agglutination, which, owing to its speed and simplicity, is very beneficial for diagnostic detection of Y. pestis antigen. Recombinant 4D5 scFv antibodies designed and conjugated with polymer particles containing quantum dots provide multipoint highly specific binding with cancer marker HER2/neu on the surface of SKOV-3 cell.

DETECTION OF TOXOPLASMA GONDII IN SHEEP AND HUMAN IN HAMA\ SYRIA

A total of 291 serological samples of Ewassi sheep were collected from different regions in Hama Mhafazat, in order to detect the morbidity rate of Toxoplasma gondii. The reported morbidity rate was 56.01%. Besides, 72 serological samples were collected from women for the same purpose, and the prevalence of infection was reported as 68.05%. Latex rapid agglutination and Latex Plate- agglutination tests were used in the examination of samples. The Positive titers were started from titer 1/64 in sheep and cats and from 1/32 in human population. The relationship between morbidity rate and age of sheep categories was studied. It was reported that morbidity rate in ages between 1-3 months was 36.95%; in ages between 3-6 months was 49.12%, and in ages between 6-9 months old was 62.5%. In sheep the infection was characterized in ages greater than one year, generally was 63.09%. Experimental infection of mice using 50 brain sheep samples for one month revealed that 4 brains, (8%) were positive for Toxoplasma-cyst.

Antibiotic-resistant &lt;i&gt;Staphylococcus aureus&lt;/i&gt; isolated from milk in the Mafikeng Area, North West province, South Africa

The aim of this study was to isolate Staphylococcus aureus from samples of cow’s milk obtained from different farm settings and to determine their antibiotic susceptibility patterns. Gram staining, oxidase, catalase, DNase, haemolysis and the MASTASTAPH TM rapid agglutination tests were employed for bacterial identification. A total of 28 milk samples were collected and screened for the presence of S. aureus . All the samples were contaminated with S. aureus . A total of 240 S. aureus isolates were obtained during this study. The levels of contamination with S. aureus were higher in milk obtained from the communal farms in Lokaleng and Mogosane (24.6% and 35.4%, respectively)compared to the commercial farms in Rooigrond and Molelwane (17.9% and 22.1%, respectively). A large percentage of the S. aureus isolates (39%–100%) from both communal farms was resistant to methicillin (MT), ampicillin (AP), penicillin G (PG), sulphamethoxazole (Smx), oxytetracycline (OT), erythromycin (E), nitrofurantoin (NI) and streptomycin (S), but not vancomycin (V). An even higher percentage (64.2% – 100%) of the isolates from both commercial farms was resistant to sulphamethoxazole and nitrofurantoin. A comparably smaller percentage (3.4% – 4.7%) of the isolates from both communal farms was resistant to vancomycin, but all isolates from commercial farm milk were susceptible to this drug. The predominant multiple antibiotic resistant phenotypes for isolates from the commercial farms were AP-Smx-NI and MT-AP-PG-OT-Smx-NI for Rooigrond and Molelwane farms, respectively, while those for isolates from the communal farms were MT-AP-PG-Smx-E-NI-S and MT-AP-PG-OT-Smx-NI-S for Lokaleng and Mogosane, respectively. When comparing the percentage of antibiotic resistance, a significant positive correlation was observed between the isolates from the commercial farms ( r = 0.966, p S. aureus is normally resident in humans, therefore, the S. aureus present in the cows may have resulted from transmission between the two species, emphasising the need to improve sanitary conditions in the milking environment.

ESTABELECIMENTO DE UM PROTOCOLO DE SOROAGLUTINAÇÃO RÁPIDA (SAR) PARA DETECÇÃO DE ANTICORPOS PARA Salmonella Typhimurium EM SUÍNOS

Salmonella Typhimurium is an important agent isolated from cases of human food poisoning in Brazil. Animal-origin products are the main source of infection, and pork has been implicated in Salmonella transmission to humans. Programs to salmonella monitoring in swine have been carried out, using bacteriological and serological tests like ELISA. However, these tests are time-consuming and expensive. The rapid agglutination test (RAT) is cheaper, faster and easier. Aiming to standardize RAT to detect anti-salmonella antibodies in swine serum, 60 samples of swine serum were tested. They had previously displayed positive (30) or negative (30) results in the ELISA test. The results showed that RAT had sensitivity, specificity, predictive positive value and predictive negative value equal to 96.7% when non-diluted serum was tested. Thus, this test can be applied to detect antibodies against S. Typhimurium in swine serum. KEYWORDS: Antibody, diagnosis, pig, rapid agglutination test, Salmonella Typhimurium.

Seroprevalence of Salmonella and Mycoplasma gallisepticum infection in chickens in Rajshahi and surrounding districts of Bangladesh

A serological survey on the prevalence of antibodies against Salmonella and Mycoplasma gallisepticum (MG) was carried out in layer chickens in Rajshahi and surrounding districts of Bangladesh. A total of 605 sera samples were examined by rapid plate agglutination (RPA) test using commercial Salmonella and MG antigens to determine the Salmonella and MG specific antibodies. Out of 605 sera samples 14.05% had single Salmonella, 45.12% had single MG and 11.24% had their concurrent infection. Prevalence of Salmonella was recorded the highest (37.60%) in adult compared to young (16.66%). On the contrary, MG and their concurrent infections were recorded the highest (71.66% and 13.33%) in young compared to adult (50.40% and 10.40%). The prevalence of Salmonella, MG and their concurrent infections were recorded the highest (34.28%, 68.57% and 17.14%) in large flocks compared to small flocks (21.25%, 50% and 8.75%). The prevalence of Salmonella infection was the highest (30.37%) in summer followed by winter (23.69%), rainy (25%) and autumn (23.33%). The prevalence of MG infection was the highest (61.58%) in winter followed by autumn (56.88%), rainy (55%) and summer (49.63%). Whereas, their concurrent infection was the highest (12.11%) in winter followed by summer (11.85%), rainy (10.83%) and autumn (10%).

Meningitis Dipstick Rapid Test: Evaluating Diagnostic Performance during an Urban Neisseria meningitidis Serogroup A Outbreak, Burkina Faso, 2007

Meningococcal meningitis outbreaks occur every year during the dry season in the “meningitis belt” of sub-Saharan Africa. Identification of the causative strain is crucial before launching mass vaccination campaigns, to assure use of the correct vaccine. Rapid agglutination (latex) tests are most commonly available in district-level laboratories at the beginning of the epidemic season; limitations include a short shelf-life and the need for refrigeration and good technical skills. Recently, a new dipstick rapid diagnostic test (RDT) was developed to identify and differentiate disease caused by meningococcal serogroups A, W135, C and Y. We evaluated the diagnostic performance of this dipstick RDT during an urban outbreak of meningitis caused by N. meningitidis serogroup A in Ouagadougou, Burkina Faso; first against an in-country reference standard of culture and/or multiplex PCR; and second against culture and/or a highly sensitive nested PCR technique performed in Oslo, Norway. We included 267 patients with suspected acute bacterial meningitis. Using the in-country reference standard, 50 samples (19%) were positive. Dipstick RDT sensitivity (N = 265) was 70% (95%CI 55–82) and specificity 97% (95%CI 93–99). Using culture and/or nested PCR, 126/259 (49%) samples were positive; dipstick RDT sensitivity (N = 257) was 32% (95%CI 24–41), and specificity was 99% (95%CI 95–100). We found dipstick RDT sensitivity lower than values reported from (i) assessments under ideal laboratory conditions (>90%), and (ii) a prior field evaluation in Niger [89% (95%CI 80–95)]. Specificity, however, was similar to (i), and higher than (ii) [62% (95%CI 48–75)]. At this stage in development, therefore, other tests (e.g., latex) might be preferred for use in peripheral health centres. We highlight the value of field evaluations for new diagnostic tests, and note relatively low sensitivity of a reference standard using multiplex vs. nested PCR. Although the former is the current standard for bacterial meningitis surveillance in the meningitis belt, nested PCR performed in a certified laboratory should be used as an absolute reference when evaluating new diagnostic tests.

Isolation of Salmonella enterica Serovar Enteritidis in Blue-Fronted Amazon Parrot (Amazona aestiva)

Avian salmonellosis is a disease caused by bacteria of the genus Salmonella that can cause three distinct diseases in birds: pullorum diseases, fowl typhoid, and paratyphoid infection. Various wildlife species are susceptible to infections by Salmonella, regardless of whether they live in captivity or freely in the wild. The present study verified the presence of Salmonella enterica serovar Enteritidis in three captive specimens of Amazona aestiva. The study involved a total of 103 birds undergoing rehabilitation to prepare for living in the wild, after having been captured from animal traffickers and delivered to the Centrofauna Project of the Floravida Institute in Sao Paulo, Brazil. This is the first report of Salmonella Enteritidis isolation in A. aestiva that originated from capture associated with animal trafficking; Salmonella was detected during the study by the serologic method of rapid serum agglutination on a plate with bacterial isolate. The antimicrobial profile exam of the isolated samples demonstrated sensitivity to ampicillin, cefaclor, ciprofloxacin, and cloranfenicol. The three samples also presented resistance to more than four antibiotics. The presence of the genes invA and spvC was verified by PCR technique and was associated with virulence and absence of class 1 integron, a gene related to antimicrobial resistance. The commercial antigen for pullorum disease was shown to be a useful tool for rapid detection in the screening of Salmonella of serogroup D1 in Psittaciformes. New studies on Salmonella carriage in birds involved in trafficking must be performed to better understand their participation in the epidemiologic cycle of salmonellosis in humans and other animals.

Prevalence and correlates of HIV and syphilis infections among men who have sex with men in seven provinces in China with historically low HIV prevalence.

To assess HIV and syphilis infection among men who have sex with men (MSM) in Chinese regions with historically low HIV prevalence. A cross-sectional survey was conducted in 20 cities or districts across 4 provinces, 2 autonomous regions, and a municipality. Socioeconomic/behavioral risk factors were measured. Blood samples were tested for HIV via enzyme-linked immunosorbent assay and Western blot confirmation and syphilis via rapid plasma reagin and passive particle agglutination test for detection of Treponema pallidum Antibodies. Of 4983 MSM participants, 2.9% were HIV infected (range: 0%-15.1%) and 9.8% were syphilis infected (1.3%-29.3%). Syphilis infection was associated with older age [adjusted odds ratio (AOR) = 2.9; 95% confidence interval (CI): 2.0 to 4.3], not being married or cohabiting (AOR = 0.7; 95% CI: 0.5-0.9), less education (AOR = 1.3; 95% CI: 1.0 to 1.7), inconsistent condom use during anal sex with men (AOR = 1.4; 95% CI: 1.0 to 1.8), and living in inner Mongolia (AOR = 23.9; 95% CI: 9.7 to 58.6), Jilin (AOR = 7.9; 95% CI: 3.4 to 18.3), Heilongjiang (AOR = 7.1; 95% CI: 3.1 to 16.6), Liaoning (AOR = 6.1; 95% CI: 2.6 to 14.2), or Chongqing (AOR = 5.9; 95% CI: 2.5 to 13.9). HIV infection was associated with older age (AOR = 3.7; 95% CI: 2.0 to 6.7), less education (AOR = 2.9; 95% CI: 1.8 to 4.7), inconsistent condom use during anal sex with men (AOR = 1.9; 95% CI: 1.2 to 3.2), syphilis infection (AOR = 2.1; 95% CI: 1.3 to 3.4), and living either in Liaoning (AOR = 8.2; 95% CI: 1.1 to 61.4) or Chongqing (AOR = 57.2; 95% CI: 7.9 to 414.4). HIV and syphilis have reached alarming rates in China's MSM population, thus, appropriate responses are urgently needed.

Isolation of Brucella canis from a human sharing her house with infected dogs.

Objective: To report a case of bacteremia in a woman living in close contact with dogs infected with Brucella canis. Methods: Previous written consent of the woman a blood sample was taken to perform rapid agglutination test, immunofluorescence, and hemoculture. Results: Brucella canis was isolated and the isolation was confirmed by the Instituto Nacional ANLIS «Carlos G. Malbran» from Argentina. The woman did not present any symptoms associated with brucellosis. Conclusion: There is a risk of contamination by B. canis for people keeping tight contact with infected dogs.

Field Evaluation of Two Rapid Diagnostic Tests for Neisseria meningitidis Serogroup A during the 2006 Outbreak in Niger

The Pastorex® (BioRad) rapid agglutination test is one of the main rapid diagnostic tests (RDTs) for meningococcal disease currently in use in the “meningitis belt”. Earlier evaluations, performed after heating and centrifugation of cerebrospinal fluid (CSF) samples, under good laboratory conditions, showed high sensitivity and specificity. However, during an epidemic, the test may be used without prior sample preparation. Recently a new, easy-to-use dipstick RDT for meningococcal disease detection on CSF was developed by the Centre de Recherche Médicale et Sanitaire in Niger and the Pasteur Institute in France. We estimate diagnostic accuracy in the field during the 2006 outbreak of Neisseria meningitidis serogroup A in Maradi, Niger, for the dipstick RDT and Pastorex® on unprepared CSF, (a) by comparing each test's sensitivity and specificity with previously reported values; and (b) by comparing results for each test on paired samples, using McNemar's test. We also (c) estimate diagnostic accuracy of the dipstick RDT on diluted whole blood. We tested unprepared CSF and diluted whole blood from 126 patients with suspected meningococcal disease presenting at four health posts. (a) Pastorex® sensitivity (69%; 95%CI 57–79) was significantly lower than found previously for prepared CSF samples [87% (81–91); or 88% (85–91)], as was specificity [81% (95%CI 68–91) vs 93% (90–95); or 93% (87–96)]. Sensitivity of the dipstick RDT [89% (95%CI 80–95)] was similar to previously reported values for ideal laboratory conditions [89% (84–93) and 94% (90–96)]. Specificity, at 62% (95%CI 48–75), was significantly lower than found previously [94% (92–96) and 97% (94–99)]. (b) McNemar's test for the dipstick RDT vs Pastorex® was statistically significant (p<0.001). (c) The dipstick RDT did not perform satisfactorily on diluted whole blood (sensitivity 73%; specificity 57%).Sensitivity and specificity of Pastorex® without prior CSF preparation were poorer than previously reported results from prepared samples; therefore we caution against using this test during an epidemic if sample preparation is not possible. For the dipstick RDT, sensitivity was similar to, while specificity was not as high as previously reported during a more stable context. Further studies are needed to evaluate its field performance, especially for different populations and other serogroups.

Evaluation and Importance of Selected Microbiological Methods in the Diagnosis of Human Brucellosis

Brucellosis is an important public health problem in Bosnia and Herzegovina. The diagnosis of brucellosis in the country without any experiences with this kind of infection may be very difficult. The aim of this study was to evaluate diagnostic methods: Rose Bengal test, blood cultures and ELISA IgM and IgG in the patients with brucellosis. The study included 91 brucellosis patients in the period 2004 to 2007. All the patients were treated at the Clinic for Infectious Diseases, University of Sarajevo Clinics Centre. Blood cultures were positive in 28/91 (30, 8%) patients. This method often needs a long period of incubation and specimens need to be obtained early. These limitations make serology the most useful tool for the laboratory diagnosis of Brucella infection. Rose Bengal is a rapid plate agglutination test, very sensitive irrespective of the stage of the disease. In our study, Rose Bengal test was positive in all patients 91/91 (100, 0%). Brucella IgM antibodies with ELISA were positive in 59/91 (64, 8%). Brucella IgG antibodies with ELISA were positive in 51/91 (56%). In order to determine the diagnostic value of the different tests, we compared the sensitivity among test-methods: Rose Bengal test-100.0%, blood culture-30.8%, ELISA IgM-64.8% and ELISA IgG-56.1%. Sensitivity of test methods was different in the different stages of illness. It is necessary to use combination of different tests such are blood culture, Rose Bengal test and ELISA in order to ensure the diagnosis. Rose Bengal test is excellent for the screening. Blood culture is a method of choice for the diagnosis acute infection. ELISA is a very good method for the diagnostic chronic disease and relapse.

Development of Rapid Agglutination Test to Detect Chicken Marek Antibody

To our knowlege, there is no rapid agglutination test to detect antibodies to viruses which might be due to the small dimension of viral particles. Through complex formation of Staphylococcus aureus bearing protein A-rabbit IgG-anti IgY-IgY anti Marek virus, agglutination of antibodies to viruses can be achieved and visualized. To design the prototype of the test, the bacterial cells of Staphylococcus aureus were coupled to a complex compound consisting of IgG-IgY-Marek antigen. This protocol was able to detect clearly the presence of Marek antibody in chicken sera, showing the rapid, clear and distinct agglutination reaction on the glass objects. No agglutination reaction was observed in the reaction of specified pathogen free chicken sera with this prototype showing the specificity of the test. This finding demonstrates a novel rapid agglutination which can be used for the detection of antibodies to various agents.