In vitro study of immunocompatibility of humanized genetically modified pig erythrocytes with human serum / 器官移植

<b>Objective</b> To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. <b>Methods</b> The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. <b>Results</b> No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all <i>P</i><0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both <i>P</i><0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all <i>P</i><0.05). <b>Conclusions</b> The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.

Evaluation of diagnostic accuracy of latex agglutination test and E-test for the detection of methicillin-resistant Staphylococcus aureus

Background: This study was undertaken to compare the phenotypic methods of latex agglutination test and e-test with polymerase chain reaction for the detection of methicillin-resistant Staphylococcus aureus. Methods: Two hundred pus samples obtained from different clinical disciplines were subjected to the latex agglutination test and minimum inhibitory concentration by e-test (Oxacillin and Vancomycin) as per the standard guidelines. The comparison was made with polymerase chain reaction as the reference test. The diagnostic accuracy of each method was reported in terms of sensitivity, specificity, positive predictive value, and negative predictive value. Results: The sensitivity of latex agglutination test was found to be 100% whereas e-test for Oxacillin was found to be 96.67% sensitive. Higher specificity for e-test was reported (99.41%) when compared to the latex agglutination test (97.65%). Conclusions: Latex agglutination and e-tests are tests are relatively simpler, rapid, and easy-to-perform methods when compared to polymerase chain reaction. The present study reported high sensitivity and specificity values for both the tests, and therefore supports usage of the stated methods as screening tools for methicillin-resistant S. aureus. However, more multi-centric studies are recommended to precisely determine the diagnostic accuracy of these phenotypic methods.

Estudo soro-epidemiológico da leptospirose canina de amostras coletadas em bairros residenciais de Cruz das Almas-BA, Brasil / Seroepidemiologic study of canine leptospirosis from samples collected in residensial neighbothoods in Cruz das Almas ­ BA, Brazil

A leptospirose é uma doença infectocontagiosa, causada por espiroquetas antigenicamente distintas da bactéria Leptospira interrogans. Os cães representam um importante elo na transmissão da leptospirose, visto que, aparentemente sadios, podem albergar leptospiras e eliminá-las no meio ambiente. Com o objetivo de se conhecer a prevalência sorológica anti-leptospírica em cães de Cruz das Almas - BA e os fatores de risco para essa zoonose, foram coletadas amostras de sangue de 200 cães. A prova diagnóstica utilizada foi a Soroaglutinação Microscópica, testando-se 19 sorovares. Das 200 amostras testadas, 60 foram reagentes. Os resultados demonstram que a sororeatividade dos cães representam risco de disseminação da leptospirose para outros animais e para o ser humano.

Leptospirosis is an infectious disease, caused by spirochetes antigenically distinct from the bacteria Leptospira interrogans. Dogs represent an important link in the transmission of leptospirosis, since, apparently healthy, they can house leptospires and eliminate them in the environment. In order to know the anti-leptospiric serological prevalence in dogs from Cruz das Almas - BA and the risk factors for this zoonosis, blood samples were collected from 200 dogs. The diagnostic test used was Microscopic Soroagglutination, testing 19 serovars. Of the 200 samples tested, 60 were reagents. The results demonstrate that the seroreactivity of dogs represents a risk of spreading leptospirosis to other animals and to humans.

The role of the serum tube agglutination test in the monitoring of human brucellosis: evaluation of post-treatment SAT titers

SUMMARY OBJECTIVE: Positive results of the serum tube agglutination test that persist after treatment may be interpreted by clinicians as treatment failures. Therefore, our study examined the value of serum tube agglutination test in demonstrating treatment success. METHODS: In this retrospective study conducted at a single center, the pre- and post-treatment serum tube agglutination test titers of patients diagnosed with brucellosis were compared. RESULTS: The end-of-treatment serum tube agglutination test titer was negative in 24 (18%) of 139 patients diagnosed with brucellosis. The most common complaints of the patients were fever (78.4%), chills (88.5%), sweating (84.9%), anorexia (79.1%), and arthralgia (63.3%). The rate of positive blood culture before the treatment was 68.3%. The absence of fever (p=0.005) and arthralgia (p=0.024) and the pretreatment serum tube agglutination test titer of <1/160 (p=0.014) were significant markers of serological cure. CONCLUSION: Although serum tube agglutination test is an effective and very successful test in the diagnosis of brucellosis, our study shows that serum tube agglutination test is not useful in demonstrating the treatment success of human brucellosis in the early post-treatment period.

Leptospirosis in central & eastern Uttar Pradesh, an underreported disease: A prospective cross-sectional study

Background & objective: Leptospirosis is a zoonotic disease associated with potentially fatal consequences and a grossly underreported disease in Uttar Pradesh. However, only a few studies are available which report the prevalence of leptospirosis in this State. Hence, this study was undertaken to know the status of the disease in central and eastern Uttar Pradesh. Methods: A total of 143 serum and urine samples were collected from patients with acute febrile illness from July 2017 to March 2019. All the serum samples were tested for Leptospira by rapid IgM antibody card and IgM ELISA and urine samples were tested by real-time polymerase chain reaction (RT-PCR) to detect Leptospira DNA. All positive and 10 per cent negative sera from ELISA and RT-PCR (all rapid test positive were also ELISA positive) were sent to the ICMR-Regional Medical Research Centre, Port Blair for microscopic agglutination test (MAT). Results: Thirty eight (26.6%) out of 143 samples were positive for leptospirosis either by ELISA or RT-PCR. Positive results were eight (6%) by Rapid card, 32 (22%) by IgM ELISA, 10 (7%) by MAT, 10 (7%) by RT-PCR. In MAT, the most common serovar was Lai followed by Hebdomadis, Bangkinang and Pomona. Interpretation & conclusions: Leptospirosis was found to be one of the important causes for acute febrile illness in the central and eastern parts of Uttar Pradesh. The results of the present study suggest that it is necessary to increase diagnostic facility and awareness in clinicians for the screening of leptospirosis in acutely febrile patients to decrease morbidity and mortality associated with this disease.

Hemagglutination in gill capillaries of sheepshead, Archosargus probatocephalus (Perciformes: Sparidae), infected by a myxosporidean / Hemaglutinação em capilares branquiais de sargo-de-dente, Archosargus probatocephalus (Perciformes: Sparidae) infectado por um mixosporídeo

Abstract During a survey Myxozoa, four specimens of the sheepshead (18 ± 1.5 cm and 59 ± 2.5 g) (Archosargus probatocephalus) were collected in the Ipioquinha river (Maceió/AL). Transmission electron microscopy observations revealed erythrocyte agglutinations in gill capillaries located near spherical cysts containing myxospores of the genus Henneguya. This hemagglutination partially or totally obstructed the gill capillaries. Erythrocytes occurred in close adherence to each other, with a closed intercellular space. A few lysed erythrocytes were observed among agglutinated cells. The reduced lumen of the capillaries was partially filled with amorphous dense homogenous material adhering to the erythrocytes. In addition, heterogeneous masses of irregular lower electron density were observed in the reduced channel of the capillary. The agglutinated erythrocytes appeared dense and homogenous, lacking cytoplasmic organelles. The nuclei had the appearance of normal condensed chromatin masses, generally without visible nucleoli. This occurrence of hemagglutination only in the capillaries located in close proximity to the developing myxozoan cysts suggests that parasite development may be a factor triggering erythrocyte agglutination. This is supported by previous experimental studies that showed a probable correlation between parasitic infections and hemagglutination. Nonetheless, further studies are necessary in order to better understand the physicochemical processes involved in this phenomenon.

Resumo Durante pesquisa de mixozoários foram coletados quatro espécimes do peixes sargo-de-dente (18 ± 1.5 cm e 59 ± 2.5 g) (Archosargus probatocephalus), no rio Ipioquinha (Maceió/AL). Observações por microscopia eletrônica de transmissão revelaram aglutinação de eritrócitos em capilares branquiais localizados próximos a cistos esféricos, contendo mixosporos do gênero Henneguya. Essa hemaglutinação obstruiu parcial ou totalmente os capilares branquiais. Os eritrócitos apareceram em forte aderência entre si, com espaço intercelular fechado. Foram observados poucos eritrócitos lisados entre as células aglutinadas. O lúmen reduzido dos capilares foi parcialmente preenchido com material homogêneo denso amorfo aderido aos eritrócitos, além de massas livres heterogêneas de densidade eletrônica baixa e irregular observadas no canal reduzido dos capilares. Os eritrócitos aglutinados pareciam densos e homogêneos, sem organelas citoplasmáticas. Os núcleos apareceram como massas normais de cromatina condensada, geralmente sem nucléolos visíveis. A ocorrência de hemaglutinação apenas nos capilares, localizados nas proximidades dos cistos mixozoários, sugere que o desenvolvimento parasitário pode ser um fator desencadeante da aglutinação eritrocitária. Isso é corroborado por estudos experimentais anteriores que mostraram uma provável correlação entre infecções parasitárias e hemaglutinação. No entanto, novos estudos são necessários para melhor compreender os processos físico-químicos envolvidos neste fenômeno.

Relación entre género y niveles de proteína C reactiva / Relation among gender and levels of C reactive protein

RESUMEN Fundamento: La proteína C reactiva es uno de los mejores marcadores para la valoración y seguimiento de enfermedades inflamatorias; los valores de referencias recomendados para su concentración en suero no están ajustados según género. Objetivo: Determinar si la concentración de proteína C reactiva difiere según el género. Metodología: Se realizó un estudio exploratorio en 3199 muestras de pacientes procedentes del Hospital General Provincial Camilo Cienfuegos y en un grupo control de 76 muestras de sueros de donantes de sangre para cada género, procedentes del Banco de Sangre de Sancti Spíritus. Los niveles se midieron a través un ensayo semicuantitativo de aglutinación con partículas de látex para la muestra supuestamente enferma y uno cuantitativo inmunoturbidimétrico para la supuestamente sana. Se compararon los niveles medios entre género en cada una mediante la prueba t de Student para muestras independientes. Resultados: La media de los niveles de proteína C reactiva en el género masculino y femenino de la muestra supuestamente enferma fue de 3.49 mg/L y 3.41 mg/L respectivamente. En el grupo control la comparación de medias de los niveles de proteína C reactiva entre género fue para los hombres de 1.38 mg/L y para las mujeres de 1.94 mg/L. Conclusión: No se encontraron diferencias significativas entre género en la muestra supuestamente enferma, ni en el grupo control.

ABSTRACT Background: C-reactive protein is one of the best marker for the assessment and monitoring of inflammatory diseases; the recommended reference values for its serum concentration are not gender-adjusted. Objective: To determine whether C-reactive protein concentration differs by gender. Methodology: An exploratory study was conducted on 3199 patient samples from Camilo Cienfuegos General Provincial Hospital and a control group of 76 serum samples from blood donors of each gender from the Sancti Spíritus Blood Bank. Levels were measured by a semi-quantitative latex particle agglutination assay for the presumed diseased sample and a quantitative immunoturbidimetric assay for the presumed healthy sample. Mean levels were compared between genders in each using Student's t-test for independent samples. Results: The mean of C-reactive protein levels in the male and female gender of the supposedly diseased sample was 3.49 mg/L and 3.41 mg/L respectively. In the control group the mean comparison of C-reactive protein levels between genders was 1.38 mg/L for males and 1.94 mg/L for females. Conclusion: No significant gender differences were found in the presumed ill sample, nor in the control group.

Antibodies against Leptospira spp. in bovine serum samples from several Brazilian states analyzed in the period from 2007 to 2015 / [Anticorpos contra Leptospira spp. em amostra de soro bovino de diversos estados brasileiros analisados no período entre 2007 e 2015]

Bovine leptospirosis assumes great economic importance since it affects several production aspects. Therefore, knowledge about the occurrence and distribution of this disease is fundamental to adopt the correct prevention measures. The present study aimed to evaluate the frequency of anti-Leptospira spp. antibodies in 24,483 bovine serum samples received between 2007 to 2015 from 21 Brazilian states. Of these, 8,643 (35.3%) were reagents in the microscopic agglutination test to one or more serovars of Leptospira spp. The most frequent serovars were Wolffi (61.47%), Tarassovi (9.62%) and Pomona (7.20%). Hardjo serovar presented a prevalence of 6.27%. Among the 21 states analyzed, the State of Pernambuco had the highest frequency with 88.24% and the State of São Paulo was the origin of the largest number of analyzed samples (13,838), with a frequency of 31.54% of reagents. The results demonstrate a high exposure to several serovars of Leptospira spp. in bovine species in Brazilian states, showing the importance of adopting prophylactic measures in order to reduce the risk of infection in this specie.(AU)

Com o objetivo de avaliar a frequência de anticorpos anti-Leptospira spp., foram analisadas 24.483 amostras de soro sanguíneo bovino, provenientes de 21 estados brasileiros, recebidas no período de 2007 a 2015. Destas, 8.643 (35,3%) foram reagentes no teste de soroaglutinação microscópica a uma ou mais sorovariedades de Leptospira spp., e as sorovariedades com maior frequência foram Wolffi (61,47%), Tarassovi (9,62%) e Pomona (7,20%). A sorovariedade Hardjo apresentou prevalência de 6,27%. Entre os 21 estados analisados, o estado de Pernambuco apresentou a maior frequência, com 88,24%, e o estado de São Paulo foi a origem do maior número de amostras para análise, 13.838, com frequência de 31,54% de reagentes.(AU)

Blood type incompatibility caused by cold agglutination: A study of 16 cases / 中国输血杂志

【Objective】 To investigate the effect of cold agglutination on blood group typing. 【Methods】 37℃ water bath, absorption elution test and 2-mercaptoethanol method were used to eliminate the influence of cold agglutination. Forward and reverse blood group typing, cross matching, DAT and IAT experiments were then performed on red blood cells and serum after treatment. 【Results】 Before treatment, obvious discrepancy in forward /reverse typing and nontypable cross matching in 16 blood samples were noticed due to cold agglutination. After corresponding treatments, all samples were consistent or negative in forward/reverse typing, cross matching and antibody screening. No adverse reactions to cross matching blood transfusion occurred in patients, and the increase of hemoglobin was in line with the effective standard of transfusion. 【Conclusion】 37℃ water bath, absorption elution test and 2-mercaptoethanol method can be used to eliminate the interference caused by cold agglutination to obtain correct typing results. The strong reactivity caused by cold agglutination in AIHA patients were different from other cases, which deserved our attention.

Anovel solid-phase agglutination reagent for detecting IgG irregular alloantibodies of red cell blood groups / 中国输血杂志

【Objective】 To develop a novel solid-phase agglutination reagent for detecting IgG irregular antibodies of red cell blood groups and evaluate its performance. 【Methods】 Monoclonal anti-RBC antibody was coated on the bottom of the microwell strips, then RBCs were bonded to the antibody and formed the monolayer by dispensing 100 μL RBC suspesion to microwell strips.RBC antigen membrane monolayer was formed by lysing RBC layer with ddH2O, then the drying medium was added to the strips and dried under reduce pressure in a vacuum dryer, thus the dried reagent microplate was obtained.Serial diluted solutions of polyclonal sheep anti-human globulin(IgG+ C3d/4)was used to react with IgG anti-D sensitized O group RBCs to select out the best indicator.Stability of membrane antigen was tested by detecting IgG anti-D and anti-E with the lowest titer by different batches of regeats. Sensitivity of the novel reagent, Capture-R Ready Screen and microcolumn gel card was carried out by detecting irregular antibodies with different titer.350 plasma samples were tested by the novel reagent and Capture-R Ready Screen to evaluate their detection ability. 【Results】 Anti-RBC solution with concentration of 20 μg/mL could fix the membrane monolayer very well on the bottom of microstrips. Sixteentimes dilution of polyclonal sheep anti-human globulin and anti-D sensitized RBCs were selected out as the best indicator.Antigen reactivity of dried RBC membrane was not weakened during the 6-monthstorage period.Sensitivity of the novel reagent was higher than Capture-R Ready Screen and microcolumn gel card. The positive consistence ratio of the novel reagent and Capture-R Ready Screen was 98.0%, the negative consistence ratio was 99.66%, and the total consistence ratio was 99.43%. 【Conclusion】 A novel solid-phase agglutination reagent with higher sensitivity and longer storage time has been developed successfully and it has an equal detection ability compared with Capture-R Ready Screen for detecting irregular alloantibodies of red cell blood groups.

Clinical features of six patients with neurobrucellosis / 中华神经医学杂志

Objective:To explore the clinical characteristics of neurobrucellosis to improve the diagnoses and treatments of this disease.Methods:Six inpatients diagnosed with neurobrucellosis in our hospital from Junuary 2014 to December 2019 were chosen, and their epidemiological features, clinical manifestations, laboratory tests, treatments, and prognoses were analyzed.Results:Six patients were male, 2 had epidemiological history, one was with orchitis, one was with imaging abnormalities related to cerebral small vessel diseases; 3 patients had brucella malta in the cerebrospinal fluid and one had brucella malta in the blood; cerebrospinal fluid (CSF) rose bengal plate agglutination test was positive in 2 patients, and the next-generation sequencing results of CSF showed that they were brucella nucleic acid sequences. All 6 patients had good prognosis after antibiotic therapy.Conclusion:The clinical manifestations of neurological brucellosis are diverse; the combined application of cerebrospinal fluid culture, cerebrospinal fluid brucella agglutination test, and next-generation sequencing is of great value in the diagnosis of neurobrucellosis.

Epidemiological characteristics of Leptospira spp. infection in bovine herds in Mato Grosso do Sul, Brazil / Caracterização soroepidemiológica da infecção por Leptospira spp. em rebanhos bovinos de corte do estado de Mato Grosso do Sul, Brasil

The epidemiological characteristics of bovine leptospirosis in animals and herds in Mato Grosso do Sul were investigated to determine parameters such as disease frequency and the serovars reactant in beef cattle herds. A total of 4,629 beef cattle herds were examined against 33 Leptospira spp. serovars. The serum samples were submitted to the microscopic agglutination test (MAT) for the serological diagnosis of leptospirosis. The MAT results showed that 3,814 (82.39%) of the 4,629 animals evaluated were seropositive for the bacterium, with serological reactions mainly to serogroup Sejroe, serovar Wolffi (36.49%). The observed high frequency of reactive animals demonstrates the relevance of the infection. Therefore, general and specific measures should be implemented to contain and/or prevent infection of the animals in the studied region.(AU)

Foi realizado um inquérito epidemiológico da leptospirose em bovinos de rebanhos de corte do estado de Mato Grosso do Sul, de modo a determinar a frequência e as sorovariedades reagentes. Para isso, foram examinados 4.629 bovinos de corte, com uma coleção de 33 sorovariedades de Leptospira, por meio da prova de Soroaglutinação Microscópica (MAT). Dos 4.629 animais examinados, 3.814 (82,39%) foram reagentes com reações predominates para o sorogrupo Sejroe, sorovar Wolffi (36,49%). Assim, a alta frequência de animais reagentes encontrada justifica a implantação de medidas gerais e específicas para conter e/ou prevenir a infecção nos animais dessa região.(AU)

Leptospirosis diagnosis among patients suspected of dengue fever in Brazil

The early symptoms of leptospirosis and dengue fever are difficult to distinguish and can cause diagnostic confusion. Due to the large dengue epidemics that has occurred in Brazil in recent years, it is possible that cases of leptospirosis were unreported. Therefore, we performed a retrospective study to detect leptospirosis in patients who were tested for dengue, but whose laboratory diagnoses were negative. Methods: Sera samples from 2,017 patients from 48 cities located in the central region of São Paulo state, Brazil, were studied. All samples were subjected to the microscopic agglutination test (MAT), 305 of which were taken from patients five days or less since the onset of symptoms, and were additionally subjected to real-time polymerase chain reaction (PCR). Results: The overall prevalence of leptospirosis cases was 21 (1.04%), with 20 through MAT (18 for Icterohaemorrhagiae and two for the Cynopteri serogroup) and one through PCR (amplicon sequencing compatible with Leptospira interrogans). According to previously established criteria, eight cases of leptospirosis were classified as "confirmed" and 13 as "probable". The Brazilian notification system for health surveillance had no records for 16 patients positive for leptospirosis and, thus, they were considered unreported cases. Statistical analyses revealed that the prevalence of leptospirosis was higher in men (1.56%) than in women (0.56%), and the mean age was higher in positive patients (43.7 years) than in negative ones (32.3 years). Conclusion: The results indicated that patients suspected of dengue fever had evidence of leptospirosis or Leptospira infection, and most of these cases were unreported in the Brazilian notification system. The high burden of dengue may contribute to the misdiagnosis of leptospirosis, and health professionals should increase their awareness of leptospirosis as an important differential diagnosis of patients with suspicion of dengue.(AU)

Production of a latex agglutination reagent for the rapid diagnosis of cryptococcal meningitis / Producción de un reactivo de látex para el diagnóstico rápido de criptococosismeníngea

Abstract Cryptococcosis is a fungal disease affecting more than one million people per yearworldwide. Its main etiological agents are Cryptococcus neoformans species complex and Cryp-tococcus gattii species complex. Cryptococcal meningitis (CM) is considered an AIDS-definingcondition. Rapid diagnosis by cryptococcal antigen assays, either the latex agglutination test(LA) or the lateral flow assay, is key to decreasing mortality due to cryptococcal disease. Theaim of the study was to develop a latex agglutination reagent (LA-ANLIS) for the rapid and reliable diagnosis of cryptococcosis in Argentina. This reagent will be produced in order to supplythe NMLN (National Mycology Laboratory Network). The evaluation of LA-ANLIS performanceand its comparison with the Cryptococcus Antigen Latex Agglutination Test System (LA-IMMY)(Immuno-Mycologics, Inc., USA) were conducted in 94 samples of cerebrospinal fluid. LA-ANLISand LA-IMMY compared exhibited 100% positive agreement and 97% negative agreement. LA-ANLIS showed 94% sensitivity and 97% specificity with the positive and negative predictivevalues of 94% and 97%, respectively. The LA-ANLIS is a reliable, reproducible and cost-effectivereagent, especially useful in countries where the commercial kit is not generally available andmust be obtained at a high cost. National production of reagents is the best choice for a reliableaccess to the rapid diagnosis of CM in Argentina.

Resumen La criptococosis es una enfermedad fúngica que afecta a más de un millón de personas por año en todo el mundo. Los principales agentes etiológicos pertenecen a los complejos de especies Cryptococcus neoformans y Cryptococcus gattii. La criptococosis meníngea (CM) se considera una enfermedad marcadora de sida. El diagnóstico rápido de esta enfermedad a través de la detección del antígeno de Cryptococcus, ya sea por aglutinación en partículas de látex o por inmunocromatografía, es clave para disminuir la mortalidad. El objetivo del presente estudio fue desarrollar un reactivo de aglutinación en partículas de látex para el diagnóstico rápido y certero de la CM en Argentina. Este reactivo (denominado en adelante LA-ANLIS) será producido para abastecer a la Red Nacional de Laboratorios de Micología. Se evaluó el desempeno del reactivo LA-ANLIS, y se realizó una comparación con el reactivo comercial Immuno-Mycologics, Inc. (en adelante, LA-IMMY) utilizando 94 muestras de líquido cefalorraquídeo. Hubo un 100% de acuerdo positivo y un 97% de acuerdo negativo entre los resultados obtenidos con los reactivos LA-ANLIS y LA-IMMY. El reactivo LA-ANLIS mostró una sensibilidad del 94% y una especificidad del 97%; los valores predictivos positivo y negativo fueron del 94 y del 97%, respectivamente. Se concluye que el LA-ANLIS es un reactivo confiable y rentable, que arroja resultados reproducibles, por lo que es especialmente útil en países donde los reactivos comerciales generalmente no están disponibles o sus costos son elevados. La producción nacional de reactivos es la mejor opción para asegurar el acceso de todos los hospitales al diagnóstico rápido de la CM en Argentina.

Síndrome vulvo vaginal gingival / Vulvo vaginal gingival syndrome

RESUMEN El liquen plano es una enfermedad inflamatoria crónica mucocutánea, que puedeafectar piel, pelo, uñas y mucosas. La afectación concomitante de la mucosa oral y vaginal, conocida como síndrome vulvovaginal-gingival del liquen plano, es una variante infrecuente y grave del liquen plano caracterizada por erosiones o descamación de las mucosas: vulvar, vaginal ygingivo-oral, con predilección por la formación de cicatrices y estenosis. De curso evolutivo benigno, aunque en ocasiones puede sufrir una degeneración maligna. Presentamos el caso de una paciente de sexo femenino de 68 años con diagnóstico de Síndrome vulvovaginal-gingival.

ABSTRACT Lichen planus is a chronic mucocutaneous inflammatory disease, which can affect the skin, hair, nails, and mucosa. Concomitant involvement of the oral and vaginal mucosa, known as vulvovaginal-gingival syndrome of lichen planus, is an infrequent and severe variant of lichen planus characterized by erosions or peeling of the mucosa: vulvar, vaginal, and oral gingival, with a predilection for formation of scars and stenosis. Of benign evolutionary course, although sometimes it can suffer a malignant degeneration. We present the case of a 68-year-old female patient diagnosed with vulvovaginal-gingival syndrome.

Detection of anti-Leptospira spp. agglutinins in captive South American river turtles, Podocnemis expansa

Leptospirosis is a zoonosis transmitted by contact with infected urine or water contaminated with the agent. Searches for Leptospira spp. in reptiles are scarce although most species have contact with aquatic environments. We evaluated the presence of anti-Leptospira spp. antibodies in Podocnemis expansa housed at the Amazonian Zoobotanical Garden, in Belém, Pará state, Brazil. We analyzed 74 serum samples through the microscopic agglutination test using 31 live antigens from different Leptospira spp. serogroups. Thirty samples (40.5%) were positive against Leptospira spp., with titrations between 100 and 3,200 for one or more serogroups. The Hebdomadis serogroup was the most prevalent, with 26 (87%) out of the 30 positive samples, followed by Djasiman, with two (7%) and Celledoni and Bataviae with one (3%) sample each. The detection of anti-Leptospira spp. agglutinins in P. expansa suggests that the aquatic environment is a transmission route for this pathogen among chelonians. (AU)

Prevalencia de leptospirosis en perros de trabajo vacunados y en población humana con riesgo ocupacional / Prevalence of leptospirosis in vaccinated working dogs and humans with occupational risk

Introducción. Los perros de trabajo pueden infectarse con diversas serovariedades de Leptospira que se mantienenen sus túbulos renales e intersticios por mucho tiempo, por lo que se convierten en portadores y fuentes de infección para otros huéspedes. Objetivo. Establecer la prevalencia de Leptospira spp. en perros de trabajo vacunados y en población humana con riesgo ocupacional de seis unidades policiales en Colombia. Materiales y métodos. Mediante tres instrumentos estructurados, se evaluaron 92 perros de trabajo con inmunización previa contra Leptospira spp. (65 machos y 27 hembras) y 69 personas de seis unidades policiales de los municipios de Manizales, Pereira, Armenia, Ibagué, Tuluá y Cali. Se obtuvieron muestras sanguíneas de las personas y de los perros, las cuales se evaluaron mediante la prueba de microaglutinación (Microscopic Agglutination Test, MAT) en 24 serogrupos. Se hizo un examen clínico de los perros y se obtuvieron muestras de orina para urocultivo. Resultados. La seroprevalencia de leptospirosis en las personas fue de 2,9 % (n=2) y en los perros de 57,61 % (n=53). Los serogrupos más prevalentes en los perros fueron Leptospira canicola y L. panama. El urocultivo fue positivo en 58,7 % (54/92) de las muestras y se encontró asociación estadísticamente significativa entre la edad de los perros (≥10 años; p=0,043) y la ubicación de la unidad policial (p=0,016). Conclusión. Las características epidemiológicas de la leptospirosis en los perros sugieren una presentación endémica de la infección. Se requieren algoritmos diagnósticos sensibles y específicos para investigar la leptospirosis canina y diferenciar los anticuerpos vacunales de la infección natural.

Introduction: Working dogs have been identified as a risk group for developing leptospirosis because they can be infected by Leptospira spp., which can be kept in the renal tubules and interstices for a long time, making them carriers and sources of infection for other hosts, including humans. Objective: To establish the prevalence of Leptospira spp. in vaccinated working dogs and in the occupationally exposed population in six police units in Colombia. Materials and methods: A total of 92 vaccinated dogs (65 males and 27 females) and 69 people from six police units in the municipalities of Manizales, Pereira, Armenia, Ibagué, Tuluá, and Cali were tested. Three structured instruments were applied and blood samples were obtained from people and dogs, which were processed with the microagglutination test (MAT) in 24 serogroups. A clinical examination of the dogs was performed and urine samples were obtained for urine cultures. Results: The seroprevalence of human leptospirosis was 2.9% (n=2) and in dogs, it was 57.61% (n=53). Among the dogs, serogroups L. canicola and L. panama were the most prevalent. Urine cultures were positive in 58.7% (54/92) of the samples. A statistically significant association was found between the age of the dogs (≥10 years; p=0.043) and the location of the police unit (p=0.016) with the urine culture. Conclusion: The epidemiological characteristics of leptospirosis in dogs suggest an endemic presentation of the infection. There is an urgent need to improve current diagnostic methods to investigate canine leptospirosis and differentiate between vaccine and natural infection antibodies.

Serologic Evidence of Leptospirosis in Patients Attending a Tertiary Care Hospital, Central Kerala, India- A Retrospective Study.

Leptospirosis is a zoonotic disease seen all over the world caused by pathogenic members of the genus Leptospira. Humans are infected by direct or indirect contact with an environment contaminated by urine of infected animals such as rodents, cattle and dogs. As the clinical manifestations of leptospirosis are nonspecific, laboratory diagnostic methods are needed for definitive diagnosis.METHODSThis is a retrospective study conducted in the Department of Microbiology, Government Medical College, Kottayam, Kerala, in patients with clinical features of acute febrile illness from January 2015 to December 2018. A total of 2941 blood samples were received for leptospira serology. They were subjected to specific anti leptospira IgM ELISA by Panbio Diagnostics, Brisbane, Australia. In the year 2018, 43 samples received during flood period were subjected to PCR, in addition to antileptospira IgM ELISA. For PCR, blood in EDTA, CSF and urine were sent to Manipal Centre for Virus Research, Karnataka. Twenty-five blood samples were sent for Microscopic Agglutination Test to Department of Veterinary Microbiology, College of Veterinary and Animal sciences, Mannuthy, India. Modified Faine’s criteria were assessed based on clinical, epidemiological and laboratory findings.RESULTSAmong 2941 cases of acute febrile illness, 777 (26.4 %) cases were serologically positive (IgM ELISA) for leptospirosis. Among 777 cases, maximum cases 481 (61.9 %) were seen during the year 2018, followed by 106 (13.64 %) cases during the year 2016, 104 cases (13.38 %) during the year 2015 and 86 (11.06 %) cases during the year 2017. Maximum cases were seen during the month of August, September, and October. Among 43 cases during flood 2018, 35 were (81.4 %) males and 8 (18.6%) were females. Maximum cases were seen in the age group of 41-50 years. Among the 43 cases, fever was present in all the 43 (100%) cases followed by myalgia in 36 (83.7 %) cases. Headache was seen in 22 (51.16%) cases, jaundice in 16 (37.2%) cases, cough and breathlessness in 15 (34.9%) cases, conjunctival suffusion in 13 (30.2% ), oliguria in 13 (30.2%) cases, vomiting in 11 (25.6 %) cases, abdominal pain in 7 (16.3%) cases, diarrhoea in 4 (9.3% ) cases, altered sensorium in 3 (6.9%) cases. Among complications, acute kidney injury was seen in 13 (30.2%) cases, followed by respiratory injury (ARDS) requiring mechanical ventilation in 10 (23.3%) cases and myocarditis in 4 (9.3%) cases.CONCLUSIONSLeptospirosis is a zoonotic disease of protean clinical manifestations with difficulty in diagnosis. Confirmation by diagnostic test is essential so that early diagnosis helps in early treatment and prevention of complications in turn reducing mortality.

Leptospirosis in slaughtered cows in the Triangulo Mineiro, Minas Gerais: prevalence, serological profile and renal lesions / Leptospirose em vacas abatidas no Triângulo Mineiro, Minas Gerais: prevalência, perfil sorológico e lesões renais

Leptospirosis is a zoonosis that affects several species of domestic and wild animals and is an important cause of economic losses in cattle in Brazil. In this study, we determined the prevalence of bovine leptospirosis in the Triângulo Mineiro region, Minas Gerais, Brazil, identified the most frequent serovars of Leptospira interrogans, and examine the renal pathological changes associated with the disease. Samples of blood serum and kidney fragments from 100 bovine females were collected in cattle abattoir. In the serological investigation 48% of the cows were positive. The serovars for which there were more reactive animals were Wolffi (24%), Hardjo (21%) and Hebdomadis (18%). Among the positive samples, 14/48 showedantibody titers greater than 1:100, and 70.83% of the seropositive animals responded to more than one Leptospira interrogans serovar. Only one farm did not have seropositive cows and in nine farms studied, six (66.66%) presented seropositive animals to the Hebdomadis serovar. At the histopathological examination, the most frequent microscopic lesions in positive animals were hyalinization (81.25%), congestion (81.25%) and hydropic degeneration (70.83%). However, these histopathological alterations were also found in kidneys of animals negative to serology, such as hyalinization (80.77%), congestion (48.07%) and hydropic degeneration (55.77%) and these findings are unrelated to positivity. Histopathological examination of the kidneys is not indicated to replace the serological diagnosis of leptospirosis, and may be used only as a complementary examination. Despite the low frequency of seropositive animals in the Triângulo Mineiro region, the disease is present in a large number of farms. Noteworthy is the high frequency of serovar Hebdomadis and it can be considered an emerging serovar in the region. The evaluation of the frequency of this serovar in other regions becomes important, and once verified should result in the recommendation of the inclusion of this serovar in the leptospirosis control.

A leptospirose é uma zoonose que afeta várias espécies de animais domésticos e silvestres e importante causa de prejuízos à bovinocultura nacional. Objetivou-se determinar a prevalência da leptospirose e identificar os sorovares de Leptospira interrogans mais freqüentes na região do Triângulo Mineiro, além de avaliar microscopicamente as lesões renais e correlacionar estas lesões com sorovares específicos. Foram coletadas amostras de soro sanguíneo e fragmentos de rim de 100 fêmeas bovinas em abatedouro e, destas 100 amostras, 48% foram positivas. Os sorovares para os quais houve mais animais reagentes foram Wolffi (24%), hardjo (21%) e Hebdomadis (18%). Dentre as amostras positivas, 14/48 apresentaram títulos de anticorpos aglutinantes superiores a 1:100 e notou-se que 70,83% dos animais soropositivos reagiram a mais de um sorovar de Leptospira interrogans. Somente uma propriedade não possuía vacas sororreagentes para leptospirose e das nove propriedades estudadas, seis (66,66%) apresentaram animais soropositivos ao sorovar Hebdomadis. Ao exame histopatológico, as alterações microscópicas mais encontradas em animais positivos foram hialinização (81,25%), congestão (81,25%) e degeneração hidrópica (70,83%). Porém, essas alterações histopatológicas também foram encontradas em rins de animais negativos à sorologia, como hialinização (80,77%), congestão (48,07%) e degeneração hidrópica (55,77%) e estes achados não apresentaram correlação com positividade. O exame histopatológico dos rins não é indicado para substituir o diagnóstico sorológico da leptospirose, podendo ser utilizado somente como um exame complementar. Apesar da frequência de animais sororreagentes ser baixa na região do Triângulo Mineiro, a doença está presente em um grande número depropriedades. Chama a atenção a alta frequência do sorovar Hebdomadis, podendo ser considerado um sorovar emergente na região. A avaliação da frequência deste sorovar em outras regiões se torna importante, pois uma vez verificada, deve resultar na recomendação da inclusão deste sorovar no controle da leptospirose.

Serological Evaluation of Clinically Suspected Leptospirosis Cases in a Tertiary Care Hospital

Leptospirosis is a zoonotic infection caused by the spirochaete Leptospira, which has worldwide distribution. Laboratory diagnosis is routinely performed by serological tests like dipstick assay, lateral flow assay and latex agglutination which are rapid tests recommended for screening the disease. Microscopic Agglutination Test (MAT) is the standard test for serological diagnosis of leptospiral infection which is not included in the test panel in most of the peripheral laboratories as the procedure is laborious and it requires to maintain live leptospira. Therefore, one of the rapid tests is routinely employed for demonstration of leptospiral antibodies. Our objective was to screen the acute cases of leptospirosis by Leptochek WB IgM and PanBio IgM Elisa and compare the findings with the MAT and correlate the clinical findings with the serological tests.METHODSThis study was conducted in a tertiary care hospital in Mangalore from August 2010 to September 2013. A total of 314 cases of clinically suspected leptospirosis were included based on the Faine’s criteria. Patients’ serum was screened for leptospiral antibodies by Leptochek WB IgM, PanBio IgM ELISA and Microscopic Agglutination Test (MAT).RESULTSOut of 314 clinically suspected cases screened, seropositivity for leptospirosis by Leptochek WB-IgM, PanBio IgM ELISA and MAT was found to be 49 (15.6%), 65 (20.7%) and 78 (24.8%) respectively. Thus, an overall prevalence rate of leptospirosis was 24.8% (78/314) based on the MAT test. Sensitivity, specificity, positive predictive value and negative predictive value of Leptocheck WB IgM was 53.8%, 97%, 85.7 and 86.4% with MAT while the comparative values of PanBio-ELISA IgM with respect to MAT test was 74.5%, 97 %, 89.2% and 92% respectively. Common clinical features among MAT positive cases were fever, chills and rigors, oliguria, vomiting, jaundice and headache.CONCLUSIONSMAT is a standard serological test for Leptospirosis. This test is not always available for peripheral health centres, as the test is time consuming and cumbersome. Thus, screening tests are now being employed for screening the patients. Rapid tests like Leptocheck -WB can be supplemented with an ELISA test for screening of clinically suspected cases of Leptospirosis and later confirmed with the MAT at reference centres.