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PubMed2001

Development and validation of an ELISA for the detection of leptospire-specific antibodies in rodents.

An enzyme-linked immunosorbent assay (ELISA) for the detection of IgG antibodies in rodents was developed and validated with the microscopic agglutination test (MAT) and leptospiral cultures. Sonicated antigen from cultures of serovars tarassovi and pyrogenes was used. As conjugate, a combination of anti-rat and anti-hamster IgG labeled with peroxidase was used. The optimal cut-off point was determined by plotting the sensitivity and specificity for various cut-off point values by means of receiver operating characteristic (ROC) curve. Concordance between ELISA and each of the MAT titers was measured by kappa (kappa). Proportions of positive results were compared by means of McNemar's test. Total 214 rodents were trapped, but only 117 could be processed by the three techniques (culture, ELISA, MAT; 1:20, 1:40, 1:50) and used for statistical analysis. Although, MAT titers in rodents infected with the serogroup Ballum tended to be lower than those infected with the serogroup Icterohaemorrhagiae, all (20/20) were ELISA-positive and almost all (19/20) were MAT-positive.The percentage of positive results obtained by ELISA, 47.0% exceeded significantly the 40.2% obtained by MAT (1:50). Difference between ELISA and MAT (1:40) was not significant and no differences were observed between ELISA and MAT (1:20). Agreement, specificity, sensitivity and the consequent area under the ROC curve between ELISA and MAT were higher as MAT cut-off points were lowered, being optimal at 1:20. The fact that differences between ELISA and MAT were significant at 1:50, but not at 1:40 or 1:20, supports the suggestion that lower MAT titers should be considered positive in rodents. The ELISA developed to detect leptospire-specific antibodies had optimal sensitivity and specificity in relation to MAT and it is concluded that it may constitute a very useful indicator for epidemiological purposes of past or present leptospiral infection in rodents.

Agglutination Tests↗
PubMed1991

Serologic survey for bovine pathogens in free-ranging European bison from Poland.

From 1980 to 1983, blood was taken from 60 selected European bison (Bison bonasus) in Poland. Serum samples were tested for the presence of antibodies against Brucella abortus, 14 serovars of Leptospira interrogans, Chlamydia spp., Coxiella burnetti; foot and and mouth disease virus, bovine leukemia virus and bovine herpes virus-1. In addition, an attempt was made to isolate bovine herpes virus-1 from the prepuce of selected bulls. Serological tests suggested chlamydial infection in 28 bison, subclinical Q-fever of a 2-yr-old heifer, subclinical bovine leukemia virus infection in a 12-yr-old bull and bovine herpes virus-1 infection in five bulls and three cows. Attempts to isolate bovine herpes virus-1 were not successful. These results suggest the possibility of cross transmission of several of these bovine pathogens between free-ranging bison and domestic cattle in Poland.

Agglutination Tests↗
PubMedDate not supplied

[Epidemiology of leptospirosis in the Kirov region].

The dynamics of leptospirosis morbidity in the Kirov region for many years is presented. The wide spread of leptospirosis foci, both natural and formed as the result of human activities, in the region is shown. The main role in infecting humans with leptospires belonged to the latter foci in a number of districts in the region was established. The etiological structure of leptospirosis cases among humans was represented, primarily, by leptospires of serogroup Grippotyphosa, following by serogroup Canicola and then serogroups Pomona, Tarassovi, Sejroe, Australis, Javanica. The dominating serogroups of leptospires were Grippotyphosa in wild rodents and Sejroe in agricultural animals. Persons belonging to the age group constituting the mainstay of work force (20-49 years) prevailed among the patients: town dwellers, factory and office workers. The main role in the transmission of infection to the population was played by the water factor.

Adolescent↗
PubMed2005

Short report: prospective evaluation of a multi-test strip for the diagnoses of scrub and murine typhus, leptospirosis, dengue Fever, and Salmonella typhi infection.

A multi-test strip dotblot immunoassay for the diagnosis of typhoid fever, scrub typhus, murine typhus, dengue virus infection and leptospirosis was evaluated in Thai adults presenting to hospital with acute, undifferentiated fever. The kit gave multiple positive test results in 33 of 36 patients with defined infections and was therefore not a useful admission diagnostic tool.

Adolescent↗
PubMed2001

[Notification according to the law for protection against infections].

Since January 1st 2001, the Law for Protection Against Infections decrees the duty of notification along two tracks, on the one hand by notification of infectious diseases and on the other hand by reporting the pathogens. However, the names of pathogens change due to scientific progress. Furthermore, these names include in some cases not only harmful agents of infectious diseases but also inoffensive organisms, i. e. Giardia lamblia. There are also problems in the notification of other pathogens such as Brucella sp., Campylobacter sp., Chlamydia psittaci,Clostridium botulinum, Cryptosporidium parvum, Leptospira interrogans, Listeria monocytogenes, Mycobacterium tuberculosis and similar agents, Norwalk-like viruses, Salmonella Paratyphi, Treponema pallidum, Trichinella spiralis and pathogenic serovars of Yersinia enterocolitica. Finally, the abandonment of notification for glanders, lymphogranuloma venereum, ulcus molle and variola appears to be risky, the reasons being partly unconvincing and contradictory.

Communicable Disease Control↗
PubMed1963

LEPTOSPIRAL COLONIAL MORPHOLOGY.

Stalheim, O. H. V. (University of Wisconsin, Madison) and J. B. Wilson. Leptospiral colonial morphology. J. Bacteriol. 86:482-489. 1963.-A sequence of apparent colonial types was observed with colonies of Leptospira pomona, L. canicola, L. icterohaemorrhagiae, and L. grippotyphosa in agar medium. Although some colonies of these serotypes had a different appearance initially, they eventually developed the mature or final appearance characteristic of the serotype. Colonies of freshly isolated, virulent cultures of L. pomona, L. canicola, and L. icterohaemorrhagiae were similar in appearance to colonies of avirulent strains of the same serotype. Additional studies of three stable and distinct colonial types of a laboratory strain of L. autumnalis revealed no differences in antigenicity, catalase activity, or mouse infectivity; however, differences in susceptibility to lysis by oleic acid were found. Although the colonial variants were stable during several in vitro variations, including growth in the presence of homologous antiserum and mutation to growth in a chemically characterized medium, rapid dissociation in vivo was found.

Animals↗
PubMed1984

Restriction endonuclease DNA analysis of Leptospira interrogans serovars icterohaemorrhagiae and hebdomadis.

Antigenic variants of Leptospira interrogans serovars copenhageni and hebdomadis were examined by bacterial restriction endonuclease DNA analysis with EcoRI, XhoI, SalI, BstEII, and HindIII as the digesting enzymes. The antigenic variants were stable cloned strains which had been cultivated in media containing homologous immune serum. One of the strains examined has been reported elsewhere (R. Yanagawa and J. Takashima, Infect. Immun. 10:1439-1442) as having an antigenic makeup which more closely resembles serovar kremastos than the serovar hebdomadis parent. The closely antigenically related but naturally occurring serovars icterhaemorrhagiae strain RGA and copenhageni strain M20 were examined in parallel. No differences could be shown between the hebdomadis parent and any of its mutants. Serovars copenhageni and icterohaemorrhagiae produced patterns which differed in the high-molecular-weight bands only. The Shibaura parent strain did not differ from copenhageni M20, but the Shibaura M1 strain differed from the other mutants and from icterohaemorrhagiae RGA in its high-molecular-weight bands.

DNA Restriction Enzymes↗
PubMed1986

Glycolipoprotein cytotoxin from Leptospira interrogans serovar copenhageni.

Lipopolysaccharide (LPS), glycolipoprotein (GLP) and lipid extract were prepared from Leptospira interrogans serovar copenhageni. GLP, lipid extract or purified fatty acids from lipid extract produced cytotoxic effects seen as cell enzyme leakage followed by cytotoxic death when tested in mouse fibroblast L929 cells in tissue culture. All extracts also agglutinated mouse erythrocytes but purified LPS was not cytotoxic. Neither GLP nor LPS were pyrogenic but both gelled Limulus amoebocyte lysate. Specific anti-GLP IgG neutralized the cytotoxic and haemagglutinating effect of GLP; however, at higher concentrations it enhanced the cytotoxicity of GLP and mediated lysis of the erythrocytes. A high dose of leptospires (i.e. 10(10) organisms) killed weanling mice causing pathological changes similar to those seen in acute leptospirosis. Similar results were obtained with live, dead, pathogenic and saprophytic leptospires. The results suggest that toxicity is involved in leptospiral infection and that lipid components either of whole leptospires or of a leptospiral GLP may contribute to the pathogenesis of acute leptospirosis.

Animals↗
PubMed2004

Leptospiral immunoglobulin-like proteins elicit protective immunity.

The identification of new vaccine candidates for leptospirosis is of great interest. This paper describes the identification of two homologous immunogenic proteins, LigA-m and LigB-m, of Leptospira interrogans serovar manilae strain UP-MMC-NIID, which show strong similarity with LigA found recently in L. interrogans serovar pomona. Southern blot analysis indicated the conservation of the lig genes among pathogenic leptospires. LigA-m protein was shown to be a surface-exposed lipoprotein expressed during in vitro culture. Both LigA-m and LigB-m induced protective immunity against a challenge with a homologous serovar of L. interrogans in mice. Furthermore, patients' sera infected with heterologous serovars' strains contained antibodies against LigA-m and LigB-m. Our results suggest that LigA-m and LigB-m proteins are new vaccine candidates for the prevention of leptospirosis.

Animals↗
PubMed2000

Introduction of a rapid dipstick assay for the detection of Leptospira-specific immunoglobulin m antibodies in the laboratory diagnosis of leptospirosis in a hospital in Makassar, Indonesia.

An easy, rapid and robust dipstick assay for detection of leptospira-specific immunoglobulin M (IgM) antibodies was evaluated on 403 patients admitted for hospitalization because of fever. The clinical symptoms and signs of 35 patients were consistent with leptospirosis. The final diagnosis for the remaining patients was as follows: 136 with typhoid fever, 82 with hepatitis, 74 with malaria, 48 with infections of the respiratory tract, and 20 with fever of unknown origin. The clinical diagnosis of leptospirosis was confirmed for 24 (68.6%) patients by the combined results of the microscopic agglutination test (MAT), the reference test for leptospirosis, and of IgM ELISA, a standard laboratory test for the serodiagnosis of leptospirosis. In addition, serum specimens from 8 (2.2%) patients with a final clinical diagnosis other than leptospirosis were found to be positive in MAT and/or IgM ELISA. Compared with the results of MAT and IgM ELISA a sensitivity of 91.6% and specificity of 93.6% was calculated for the dipstick assay. Most of the serum samples from the laboratory confirmed patients gave a moderate to strong staining intensity of the antigen band of the dipstick and were easy to read. The results demonstrate that the dipstick assay is convenient to use and allows the rapid and accurate confirmation of patients with clinical suspicion of leptospirosis in areas where the disease is endemic.

Antibodies, Bacterial↗
PubMed1988

Cell-mediated immune response of goats to leptospirosis.

Cell-mediated immunity to Leptospira pomona was measured in goats by the leukocyte-migration inhibition test (LMIT) and microassay for stimulation of protein synthesis (SPS) with 3H-leucine. L. pomona soluble antigen was used in both tests. The migration indices in the group of goats infected with L. pomona were significantly lower (less than .001) as compared to control. The migration zones of the unstimulated peripheral blood leukocytes in the infected group were significantly smaller (less than .001) than those of the control. The SPS test was strongly positive in test animals and was negative in control.

Animals↗
PubMed1989

Leptospirosis. Epidemiological features of a sporadic case.

Leptospirosis occurred in a 45-year-old man with presumed infection from an exposure to contaminated water at his source of employment. An intensive epidemiological investigation, including serological examination of all family members and pets and cultures on the patient and his family pets (cats and dogs), proved that the leptospiral organism was acquired by the patient's exposure to his dogs. The risk of acquiring infection from dogs that are asymptomatic and vaccinated is emphasized by this report.

Animals↗
PubMed2004

Leptospirosis misdiagnosed as pulmonary-renal syndrome.

A case of leptospirosis complicated by diffuse alveolar hemorrhage, acute renal failure, and apparent hemoglobinuria was initially diagnosed and managed as a pulmonary-renal syndrome. However, renal biopsy findings were normal. Leptospirosis may manifest with severe lung injury characterized by diffuse alveolar hemorrhage, acute respiratory distress syndrome, and acute renal failure and be accompanied by high mortality rates. Leptospirosis should be considered in patients with features of pulmonary-renal syndrome, particularly in regions known to be endemic for leptospirosis. A history of potential exposure to Leptospira is an important diagnostic clue, and empiric antimicrobial therapy should be considered.

Adrenal Cortex Hormones↗
PubMed2003

Possible scrub typhus coinfections in Thai agricultural workers hospitalized with leptospirosis.

Possible coinfections with Orientia tsutsugamushi the causative agent of scrub typhus, were prospectively evaluated in rice farmers hospitalized with leptospirosis in Northeast Thailand. Of 22 adults with leptospirosis diagnosed by the microscopic agglutination test, 9 also had serologic evidence of scrub typhus. Of 9 individuals with possible coinfections, 5 had signs or symptoms typical of scrub typhus and atypical of leptospirosis. Patients who appeared to have mixed infections had significantly higher median platelet counts and significantly lower median serum bilirubin and creatinine concentrations (P < 0.05, Mann-Whitney U test) than did individuals with leptospirosis alone. One patient with serologic evidence of scrub typhus and leptospirosis was treated only with penicillin, to which scrub typhus is not sensitive. Respiratory distress worsened during therapy, and the patient died of respiratory failure. Physicians should consider the possibility of scrub typhus infection in leptospirosis patients who respond poorly to treatment or who have atypical disease manifestations.

Adult↗
PubMed1988

Failure to demonstrate transmission of enzootic bovine leukemia virus infection from cows to sheep by use of common injection needles.

Four bovine leukemia virus (BLV)-seropositive and 2 BLV-seronegative cows were used as donors in a study to provide evidence whether IM injection with common needles is a means of spreading bovine leukemia. Sheep were used as recipients. Of the 4 BLV-seropositive cows, 2 had high virus expression (VE; 43% and 28% of their lymphocyte thin sections had associated BLV-particles), whereas the other 2 cows did not have observed VE. After each of the 4 cows was given an injection of a 5-antigen Leptospira bacterin, a BLV-seronegative sheep was immediately given an injection of the same bacterin with the same needle. None of these sheep seroconverted, nor did either of 2 sheep given only the bacterin (with a previously unused needle). Sheep inoculated IM with 0.2 ml of whole blood from both of the cows with high VE and from 1 of the 2 BLV-seropositive cows that did not have observed VE did seroconvert. In contrast, the sheep inoculated with 0.2 ml of blood from the remaining BLV-seropositive (0% VE) cow and from the 2 BLV-seronegative cows remained seronegative. These results were interpreted to indicate that the quantity of infective lymphocytes passed during injection with common needles is too small to induce infection.

Animals↗
PubMed2002

Evaluation of clinical criteria for the diagnosis of leptospirosis.

OBJECTIVES: Leptospirosis has a wide range of clinical presentation and therefore, clinical suspicion of the infection is often difficult. The objective of this study is to find out the usefulness of the clinical and epidemiological criteria in the diagnosis of leptospirosis and its comparison with microagglutination test (MAT). METHODS: A total of 118 patients with undiagnosed fever of more than seven days duration were included in the study. Their clinical presentation was scored on the basis of a clinical criteria. Sera of the patients were tested for antibodies against leptospira with the help of microagglutination test using a battery of antigens. The usefulness of the criteria was evaluated and compared with microagglutination test. RESULTS: A total of 44 out of 118 (37.28%) patients could be provisionally diagnosed as cases of leptospirosis on the basis of the clinical criteria. Eighteen of these 44 (40.9%) patients had serological evidence of leptospirosis. The criteria had a sensitivity of 81.81%, specificity of 72.91%, a positive predictive value of 40.9% and a negative predictive value of 94.59% when compared with microagglutination test. CONCLUSIONS: The criteria had a moderate sensitivity and specificity. Considering the non-specific signs and symptoms of this infection, the positive predictive value is significantly high. The criteria has a high negative predictive value and this would help the clinicians exclude the diagnosis of leptospirosis with precision.

Agglutination Tests↗