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PubMed1985

Rapid serodiagnosis of leptospirosis using the IgM-specific Dot-ELISA: comparison with the microscopic agglutination test.

The dot enzyme-linked immunosorbent assay (Dot-ELISA) was compared to the microscopic agglutination test (MA test) for the diagnosis of human leptospirosis. Of 177 sera from 68 soldiers who trained in the Republic of Panama, 102 sera were positive in the MA test and 93 of these sera were positive in the IgM-specific Dot-ELISA. Incidence of infection was 50 of 68 patients with the MA test and 48 of 68 in the IgM Dot-ELISA. Five MA test-positive sera were reactive only in the IgG-specific Dot-ELISA, suggesting previous exposure. All 21 infecting serovars of Leptospira interrogans, as determined by positive reactions in the MA test or culture of blood and urine specimens, were reactive in the Dot-ELISA. Of 75 sera negative in the MA test, 61 were nonreactive in the Dot-ELISA. However, 9 of these 14 Dot-ELISA-positive/MA test-negative sera were acute samples from patients whose later sera were MA test-positive. Positive reactions in the IgM Dot-ELISA occurred in 2 of 30 control, 4 of 10 Lyme disease, 1 of 11 relapsing fever, and 1 of 8 yaws sera; 10 syphilis patient sera were nonreactive. The IgM-specific Dot-ELISA appears to be sensitive and specific for the serodiagnosis of acute leptospirosis. In addition, this rapid test is inexpensive, simple to perform, utilizes minute volumes of killed leptospiral antigen and is easily adaptable to field use.

Agglutination Tests↗
PubMed2000

[A molecular epidemiological investigation on Leptospira interrogans serovar hebdomadis and australia in China].

OBJECTIVE: In order to confirm whether cattle serve the source of infection for patients with Leptospira interrogans serovar hebdomadis and australia. METHODS: 12 field strains of leptospira interrogans serovar hebdomadis and australia were isolated from blood samples of patients with leptospirosis and farm cattle urine. An analysis of chromosome DNA restriction endonuclease patterns (REP) and restriction fragments length polymorphism (RFLP) of 16S + 23S rRNA gene were processed by digestion of chromosome DNA using EcoR I. RESULTS: The same serovar field strains of Leptospira interrogans from blood samples of patients with leptospirosis and farm cattle urine resulted in unique restriction endonuclease patterns (REP) and ribosomal types (RT). The chromosome DNAs from field strains that belonged to different serovars of Leptospira interrogans caused different restriction endonuclease patterns and ribosomal types. CONCLUSION: In accordance with this results, we recognize that farm cattle serve the infectious source of leptospirosis serovar hebdomadis and australia thus may have realistic significance in the control of leptospirosis serovar epidemic of hebdomadis and australia in China.

Animals↗
PubMed1986

Gellan gum as a substitute for agar in leptospiral media.

An albumin polysorbate semisolid medium (Ellinghausen McCullough Johnson Harris medium) gelled with gellan gum (Gelrite; Kelco Div., Merck & Co., Inc.) compared favorably with conventional agar media for the cultivation of both pathogenic and saprophytic leptospires. The gellan gum medium supported the growth of all 18 leptospiral strains studied which included an array of serovars with various fastidious growth characteristics. Gellan gum medium was also used advantageously as a long-term maintenance medium; 9- to 12-month-old cultures still contained viable organisms. The colonial growth in gellan gum plating medium of six representative strains was consistent with previously described colonial growth on agar plating media. In addition, gellan gum medium appeared to be an excellent medium for the recovery of leptospires from the blood, liver, and kidneys of hamsters experimentally infected with a virulent Leptospira interrogans serovar bataviae strain. As few as 1 to 10 organisms in the infective tissue could be recovered in semisolid Ellinghausen McCullough Johnson Harris-gellan gum medium. The antigenicity did not appear to be affected by growth in gellan gum medium. The hamster-virulent strain of L. interrogans serovar bataviae isolated from a moribund hamster maintained its virulence after 10 sequential passages in gellan gum medium. Gellan gum medium can be a valuable adjunct to currently used cultural procedures.

Agar↗
PubMed1979

Antibodies against Leptospira biflexa serotypes patoc and são paulo in pigs: possible occurrence and importance for the intracutaneous test for leptospirosis.

Leptospirin for the diagnosis of leptospirosis by an intracutaneous test contains antigenic material from 5 pathogenic Leptospira serotypes (10). During experiments with rabbits and pigs, leptospirin was injected into 6 pigs which had been infected artificially with apathogenic Leptospira biflexa serotypes patoc and são paulo. Three out of the 6 pigs showed a positive leptospirin reaction (11). This interfering reaction in animals having been infected with apathogenic L. biflexa was the reason to investigate the occurrence of biflexa antibodies in pigs from different areas in Germany by the microscopic agglutination. None of the 854 pigs showed biflexa antibodies producing a 50% agglutination at a serum dilution of 1:100. If at all, pigs may become infected naturally by L. biflexa; this apparently seems to be a rare incident. An impairment of the diagnostic value of leptospirin by L. biflexa antibodies can be excluded.

Animals↗
PubMed1982

Leptospirosis in Rattus norvegicus and Rattus rattus in Israel.

Leptospires were isolated from 21.2% of Rattus norvegicus (the brown rat) and 9.9% of Rattus rattus trapped in a survey of urban and rural areas in Israel. Microagglutination antibodies were found in sera of 9.4 and 8.1% of these rats, respectively. The 191 strains isolated belonged to six serogroups: Leptospira icterohaemorrhagiae (169), L. bataviae (3), L. semeranga (1), L andamana (1) and L. ballum (1) from R. norvegicus and L. ballum (13) and L. hebdomadis (3) from R. rattus. No significant differences were found between the infection rate of mature male and female brown rats, but all six strains isolated from young animals were obtained from male rats. Two factors influence the rat of Leptospira infection among urban brown rats: the age of the animals and the area in which they are trapped. Adult brown rats trapped in a newly infected area showed a considerably higher level of contamination (61.8%) than those trapped in areas where the infection had persisted for many years (22.8%). Young rats showed a low degree of contamination in both the old foci (5.7%) and the new foci (2.2%). New and highly infected areas were found during this survey in Tel Aviv (Carmel market) and its neighboring cities of Holon, Ramat Gan and Givatayim.

Agglutination Tests↗
PubMed1991

Effect of vaccination with a monovalent Leptospira interrogans serovar hardjo type hardjo-bovis vaccine on type hardjo-bovis infection of cattle.

Effectiveness of 2 concentrations of a monovalent vaccine containing Leptospira interrogans serovar hardjo type hardjo-bovis was evaluated for protection of heifers from infection with type hardjo-bovis. Nine heifers were given 2 doses of low-dose vaccine (8.32 x 10(8) cells/dose); 9 heifers were given 2 doses of high-dose vaccine (8.32 x 10(9) cells/dose); and 1 steer and 1 heifer were maintained as nonvaccinated controls. Groups of vaccinated cattle were challenge-exposed with serovar hardjo type hardjo-bovis at 7 (n = 6), 11 (n = 6), or 15 (n = 6) weeks after completion of vaccination. All cattle were challenge-exposed by conjunctival instillation of 1 x 10(5) hardjo-bovis cells on 3 consecutive days. Both control and all vaccinated cattle became infected and shed serovar hardjo type hardjo-bovis in their urine. Leptospires were detected in 15 of 16 (94%) urine samples from control cattle and in 124 of 143 (87%) samples from vaccinated cattle. Leptospires were detected in kidneys of 17 of 18 vaccinated cattle and 2 of 2 control cattle and in the uterus or oviducts of 13 of 18 vaccinates and the 1 control heifer.

Agglutination Tests↗
PubMed1997

Zoonotic and viral infection in fetal loss after 12 weeks.

One hundred and thirty-six women from an urban, rural and farming community were recruited to a study of infectious causes of midtrimester miscarriage (n = 85), stillbirth (n = 32), or termination for developmental (n = 17) or chromosomal (n = 2) abnormalities. No woman had evidence of acute infection with toxoplasma, listeria, leptospira or Chlamydia psittaci (ovine enzootic abortion). One woman had midtrimester miscarriage associated with primary cytomegolovirus infection and five women had evidence of parvovirus B19 infection, although fetal infection was not proven. Zoonoses were not identified as a cause of fetal loss or malformation in this at-risk population, but parvovirus B19 was a significant cause of midtrimester fetal loss.

Abortion, Induced↗
PubMed1968

Water strains of Leptospira in the serodiagnosis of human and animal leptospirosis.

The most widely used serological reaction for the diagnosis of leptospirosis is the agglutination test. This test, however, cannot be carried out in many laboratories because special equipment and special experience are required. It is also necessary to maintain live Leptospira cultures belonging to all the serotypes present in the country where the test is made. Consequently, it would be extremely useful to be able to diagnose leptospirosis by means of a single antigen, regardless of the serotype to which the Leptospira responsible for the infection belonged. This is particularly important for countries in which the antigenic pattern of the local leptospirae is not well known and in which it would thus be necessary to use a large number of Leptospira serotypes for each test.Observations made in the last 11 years have suggested that the problem may be solved with the use of some non-pathogenic, water strains of Leptospira which seem to be agglutinated by human sera containing antibodies against pathogenic leptospirae. This paper reports the results of studies from 1960 to 1968 on the possibility of using water strains for serodiagnosis.The results over the 8-year period show that the non-pathogenic strain Patoc 1 is agglutinated by a high percentage of human sera positive for pathogenic leptospirae: these results indicate that Patoc 1 would be useful for serodiagnosis. However, a high percentage of animal sera positive for pathogenic leptospirae gave negative results with the strains Patoc 1 and Sao Paulo, and thus these strains cannot be used for serodiagnosis in animals.

Agglutination Tests↗
PubMed1993

The prevalence of small terrestrial mammals infected with tick-borne encephalitis virus and leptospirae in the foothills of the southern Bavarian forest, Germany.

In the district of Grafenau/Freyung (Bavaria, Germany), 266 specimens of small terrestrial mammals of 8 species were captured using live traps. From these mammals, Apodemus flavicollis (42.1%) and Clethrionomys glareolus (39.5%) were prevalent. All animals were tested for neutralizing antibodies to tick-borne encephalitis (TBE) virus and agglutinating antibodies to leptospirae. Seropositivity against TBE virus was 14.0% and against leptospirae 7.9%, respectively. Seropositivity to leptospirae appeared to be primarily to Leptospira grippotyphosa, less to Australis and occasionally to Javanica serovars. Only one A. flavicollis specimen was positive to both pathogens tested. The parasitocoenosis of trapped micromammals with ectoparasites consisted of 69.5% from ticks (mainly Ixodes ricinus, less from I. trianguliceps), 16.1% of mites (primarily Laelaps agilis) and 14.3% of fleas (mostly Ctenophthalmus agyrtes).

Animals↗
PubMed2006

Application of multilocus variable-number tandem-repeat analysis for molecular typing of the agent of leptospirosis.

Leptospirosis is a worldwide-distributed zoonosis, endemic in tropical areas. Epidemiologic investigations of leptospirosis still rely on tedious serological identification tests. Recently, molecular typing systems based on variable-number tandem-repeat (VNTR) analysis have been described and have been used to identify Leptospira interrogans strains. Although L. interrogans is the most common Leptospira species encountered in human infections around the world, other pathogenic species, such as Leptospira kirschneri and Leptospira borgpetersenii, are also frequently associated with human leptospirosis. In this study, we aimed to extend multilocus VNTR analysis (MLVA) identification of strains to species other than L. interrogans. We designed primers for VNTR loci found in L. interrogans, L. kirschneri, and L. borgpetersenii. The discriminatory power of the redefined primers was evaluated on collection strains and then on clinical strains. We also carried out a retrospective study on 156 strains isolated from patients and animals from New Caledonia, an area of high endemicity in the South Pacific. Our results show that this simple PCR-based MLVA typing technique is a powerful methodology for the epidemiology of leptospirosis.

Animals↗
PubMed1982

Bovine IgM and IgG response to Leptospira interrogans serovar hardjo as measured by enzyme immunoassay.

The enzyme-linked immunosorbent assay (ELISA) was used to detect specific IgG and IgG antibodies in the sera of cattle infected or immunized with Leptospira interrogans serovar hardjo. IgM appeared first but was quickly followed by IgG which persisted longer than IgM. The levels of antibody detectable by ELISA and by the microscopic agglutination test (MAT) did not correlate, suggesting that the two techniques measured different antigen--antibody systems. The transient nature of the IgM response as measured by ELISA indicates potential usefulness as a serodiagnostic test for detecting current leptospiral infections.

Agglutination Tests↗
PubMedDate not supplied

Sero surveillance of leptospirosis among sewer workers in Pune.

Leptospirosis is an important occupational disease affecting people coming in contact with animals and their discharges. The occurrence of infection in ones workplaces is linked to the environment to which the worker is exposed and the adaptability of the organism in that working environment. Rodents usually abound in underground sewers and are carriers of leptospira. The urine of rodents and other animals present in that area is likely to contaminate these sewers. Leptospira are excreted in the urine of infected animals. Thus sewer workers are at a potential risk of leptospirosis. The prevalence of leptospirosis in these workers could thus indirectly predict the presence of the disease in animals in a particular geographical niche. Total seventy-eight sewer workers from 5 different municipal wards in Pune were examined to find out the evidence of past infection with leptospira using microagglutination test (MAT). The prevalence rate was found to be 16.6%. The serovars to which antibodies were detected include autumnalis (38.4%), pyrogenes (23.0%), canicola (15.3%) and pomona (15.3%). Evidence of leptospiral infection was found to be maximum in sewer workers in the areas of the city that were infested with rodents and stray animals.

Animals↗
PubMed2005

Enzyme-linked immunosorbent assay for the detection of canine Leptospira antibodies using recombinant OmpL1 protein.

OmpL1 is a 31-kDa outer membrane protein characterized in 1993 and known to be expressed only in pathogenic Leptospira spp. Recombinant OmpL1 (GST-rOmpL1) was expressed for use as an ELISA antigen for the detection of anti-Leptospira antibodies. In immunoblot analysis, the protein reacted with sera of dogs infected with three different serotypes of Leptospira interrogans, while did not react with sera of dogs both uninfected negative controls and infected with Borrelia burgdorferi, which is closely related to Leptospira spp. Moreover, in ELISA using GST-rOmpL1, the optical density (O.D.) values from the positive controls were very high (1.125 +/- 0.549). In contrast, the O.D. values from clinically healthy dogs and dogs with diseases other than leptospirosis were very low (0.109 +/- 0.046 and 0.089 +/- 0.046, respectively). These data suggest that the detection of anti-Leptospira antibodies by ELISA using the GST-rOmpL1 protein can be applied for diagnosis of canine leptospirosis.

Animals↗
PubMed1980

Immunosuppression in bovine trypanosomiasis. The establishment of "memory" in cattle infected with T. congolense and the effect of post infection serum on in vitro (3H)-thymidine uptake by lymphocytes and on leucocyte migration.

Cattle infected with Trypanosoma congolense were intravenously immunized with Leptospira biflexa 15 days after trypanosomal infection. The primary immune response to L. biflexa was considerably reduced as compared to uninfected controls. The infected cattle mounted a secondary response when they were cured of trypanosomes by treatment with Berenil 25 days after infection and re-immunized 8 days later. The mean secondary response in these previously infected animals was lower tha, but not significantly different from that of the uninfected control animals. Serum collected 15, 20 and 25 days after infection was inhibitory to the migration of both autologous and isologous (control) peripheral blood leucocytes. The migration inhibitory activity was abolished by heating the serum at 56 degrees C for 30 minutes implying the involvement of a heat labile serum component(s). The same serum did not modify the mitogenic effect of PHA on autologous peripheral lymphocytes.

Animals↗
PubMed1966

Recent observations on leptospirosis in Northern Ireland and their bearing on current diagnostic methods.

An investigation was undertaken to assess the present importance of leptospiral infections in Northern Ireland, and in particular to look for evidence of infection by leptospiral serotypes other than L. icterohaemorrhagiae and L. canicola. Blood samples from 335 patients, sent to the laboratory for a variety of tests, were examined. After initial screening with five groups of pooled antigens, tests for leptospiral agglutinins were completed with formolized antigens prepared from 13 different serotypes. In seven patients a diagnosis of acute leptospirosis was made while nine others showed serological evidence of previous leptospiral infection. Attempts to isolate leptospirae by culture from 29 blood samples were unsuccessful. The serological results indicate that two additional leptospiral serotypes, namely L. ballum and L. bratislava, are causing human infection in Northern Ireland, and presumably also in other parts of the British Isles. Some clinical and epidemiological features associated with different types of leptospiral infection are described. It is stressed that leptospirosis is essentially a febrile illness, that meningeal symptoms are common, and that (contrary to popular belief) jaundice is by no means a constant occurrence. The implications of these findings are discussed, with special reference to the diagnosis of leptospiral infections. Laboratory diagnostic procedures are briefly reviewed, and the possible deficiencies of the agglutination test commonly used in Britain are pointed out. Some suggestions are made concerning both clinical and laboratory aspects of diagnosis, and the need for a reliable screening test for all forms of leptospiral infection is emphasized.

Adolescent↗