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Diagnosis of human leptospirosis by monoclonal antibody-based antigen detection in urine.

Hybridomas secreting specific monoclonal antibodies (MAb) to all members of the genus Leptospira (clone LF9) and those that are specific only to the pathogenic species (clones LD5 and LE1) were produced. MAb LF9, which was immunoglobulin G1 (IgG1), reacted to a 38-kDa component of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated whole-cell lysates of all Leptospira spp., while MAb LD5 and MAb LE1, which were IgG1 and IgG2a, respectively, reacted to the 35- to 36-kDa components of all serogroups of the pathogenic species of LEPTOSPIRA: The MAb LD5 was used in a dot blot-enzyme-linked immunosorbent assay (dot-ELISA) for detecting Leptospira antigen in urine samples serially collected from two groups of patients diagnosed with leptospirosis, i.e., 36 clinically diagnosed patients and 25 Leptospira culture confirmed patients. Their serum samples were tested serologically by IgM Dipstick assay, indirect immunofluorescence assay (IFA), and/or microscopic agglutination test (MAT). Urine samples of 26 patients diagnosed with other illnesses and 120 healthy individuals served as controls. For the first group of patients, who had been ill for an average of 3.4 days before hospitalization, the IgM Dipstick test, IFA, and MAT were positive for 69.4, 70.0, and 85.7% of patients, while the Leptospira antigenuria tested by the MAb-based dot-ELISA was positive for 75.0, 88.9, 97.2, 97.2, and 100% of patients on days 1, 2, 3, 7, and 14 of hospitalization, respectively. All but 1 of 11 patients whose serum samples collected on the first day of hospitalization were IgM seronegative, were positive by urine antigen test on day 1. This is strong evidence that detection of antigen in urine can provide diagnostic information that could be useful in directing early therapeutic intervention. The MAT was positive in 10 of 12 patients (83.3%) of the 25 culture-positive Leptospira patients who had been ill for an average of 5.04 days before hospitalization, and the Leptospira antigen was found in 64.0, 84.0, 96.0, 100, 100, 100, and 100% of the patients' urine samples collected on days 1, 2, 3, 4, 5, 6, and 7 of hospitalization, respectively. Leptospira antigenuria was found in 3 of the 26 patients diagnosed with other illnesses and 1 of the 120 healthy controls. The reasons for this positivity are discussed. The detection of antigen in urine by the monoclonal antibody-based dot-ELISA has high potential for rapid, sensitive, and specific diagnosis of leptospirosis at a low cost.

Animals↗

Leptospirosis in human pregnancy followed by death of the foetus.

An attack of leptospirosis due to serotype canicola in a pregnant woman was followed in the convalescent period by death of the foetus. Previous cases of this nature are reviewed. In certain areas where leptospirosis is known to exist among the animal population early recognition and treatment of human cases is advised, especially when they occur during pregnancy.

Adult↗

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↗

Q fever and leptospirosis in the dairy farming community and allied workers of Worcestershire.

Sera collected from dairy farmers, their families and farm workers, slaughtermen, artificial inseminators, veterinarians, and a group of doctors and civil servants with no direct connexion with farming were examined for evidence of Q fever or leptospirosis. One thousand and fifty-two sera examined for Q fever yielded 26% of positive results with titres of complement-fixing antibody through 1 in 4 to 1 in 128 and over. On the other hand, 876 sera examined for agglutinating antibody to various leptospirae showed only 0.5% of positive titres of 1 in 80 and over. None of the individuals with positive sera for Q fever had ever been ill with anything resembling Q fever or could remember any ill health in the past; this suggests subclinical infection. One farmer with a high titre against L. copenhageni (icterohaemorrhagiae) gave a history of a recent febrile illness very suggestive of acute leptospirosis. Infection due to leptospirae appeared to be very much less common than that due to C. burneti.

Adolescent↗

Need for vaccination of sewer workers against leptospirosis and hepatitis A.

OBJECTIVES: This study compared the prevalence of Leptospira interrogans and hepatitis A virus (HAV) antibodies in serum samples from sewer workers and controls. METHODS: A blood sample was obtained from 76 of the 101 municipal sewer workers (75%) of Quebec City and from two controls matched to each for age and sex. Testing was done for antibodies against serovars of Leptospira icterohaemorragiae, bratislava, hardjo, grippotyphosa, and kennewicki (pomona) and hepatitis A. RESULTS: Sewer workers had a greater prevalence of antibodies against leptospirosis than controls (12% v 2%, P = 0.003). In contrast, antibodies to HAV were not significantly more prevalent among workers than among controls (54% v 49%, P = 0.51). Prevalence of HAV antibodies increased significantly with age both among workers and controls (chi 2 for trend, P < 0.001). In contrast with younger workers, prevalence of HAV antibodies was greater among workers > or = 40 years than among their controls (81% v 65%, P = 0.04). CONCLUSION: Leptospirosis continues to be a problem to sewer workers but hepatitis A is apparently no longer a risk. The likely explanation is that leptospira are still abundant in the sewage system in contrast with HAV, which is only rarely to be found in sewage as a result of the generalised decrease in incidence of hepatitis A in the past three decades. The decision to vaccinate sewer workers against hepatitis A should take into account that it is impossible to avoid all contact with sewage fluid and, despite the fact that the actual incidence of hepatitis A is low, there is a real possibility of sporadic exposure during a future outbreak.

Adult↗

Leptospirosis presenting as a flaccid paraplegia.

A fatal case of leptospirosis in a 64 year old farm worker is described. The dramatic neurological presentation with a rapidly evolving flaccid paraplegia associated with biochemical evidence of renal and hepatic dysfunction is discussed. Attention is drawn to the wide range of neurological symptoms reported in leptospirosis, and to the possibility that this infectious disease may present neurologically.

Humans↗

The Jarisch-Herxheimer reaction in leptospirosis.

Three patients with leptospirosis whose condition worsened after initiation of antibiotic therapy are reported. Their clinical deterioration appeared to be due to the development of the Jarisch-Herxheimer reaction rather than to progression of their underlying infection. Relevant aspects of the management of patients with leptospirosis are discussed.

Adolescent↗

Hemolytic uremic syndrome in leptospirosis bataviae.

Hemolytic uremic syndrome was observed in a 46-year-old man who had leptospirosis. Renal failure was severe with a prolonged clinical course. Despite clinical recovery there was residual renal damage indicated by mildly elevated serum creatinine. This is the first report of hemolytic uremic syndrome in leptospirosis.

Creatinine↗

Renal involvement in leptospirosis: a pathophysiologic study.

The kidney involvement in leptospirosis appears to be a special form of acute renal failure due to a higher frequency of polyuric forms and the presence of hypokalemia with an elevated urinary fractional excretion of potassium. Using a clearance technique, we detected higher fractional urinary potassium excretion in leptospirotic guinea pigs (26.5 +/- 4.7%) than in normal animals (14.1 +/- 2.8%, p < 0.05). After blocking distal NaCl reabsorption with furosemide, it was observed that in leptospirotic animals both fractional sodium excretion (40.0 +/- 7.4%) and fractional potassium excretion (136.3 +/- 32.7%) were higher than in normal animals (20.4 +/- 3.8%, p < 0.05, and 43.6 +/- 9.0%, p < 0.05, respectively). Microperfusion studies showed that the normal and leptospirotic medullary thick ascending limb had both identical transepithelial potential difference (+3.7 +/- 0.4 vs. 3.9 +/- 0.2 mV) and relative sodium-to-chloride permeability. The same technique showed that the osmotic water permeability (Posm; 0.9 +/- 0.4 x 10(-5) cm/s.atm) and diffusional permeability (34.7 +/- 6.6 x 10(-5) cm/s) observed in the leptospirotic inner medullary collecting duct (IMCD) in the presence of vasopressin were unchanged, as was also the case for urea permeability (3.74 +/- 0.7 x 10(-5) cm/s). These data show that acute renal failure in leptospirosis is characterized by tubular changes leading to potassium secretion probably due to a decrease in proximal sodium reabsorption. Furthermore, the inability to concentrate urine evidenced by the low P(o)sm present in leptospirotic animals is due, at least in part, to IMCD resistance to vasopressin.

Animals↗

Urban rats as chronic carriers of leptospirosis: an ultrastructural investigation.

Kidneys of six wild-trapped, adult Norway rats with high agglutinating titers to Leptospira icterohaemmorhagiae were examined ultrastructurally. Although many leptospires were seen in the lumen of proximal kidney tubules, few changes in epithelial cells of this region were found. The predominant lesion in chronic rat leptospirosis was diffuse interstitial nephritis. Principal kidney lesions were proximal tubule cell basement membrane thickening with or without the presence of granular particles, and an interstitial response that varied from mixed hypercellular to chiefly plasmacytic. The association of leptospires with membrane-bound protein droplets of the proximal kidney tubules may be important in the maintenance of the chronic carrier state in rat leptospirosis.

Animals↗

Porcine leptospirosis in Iowa.

The epidemiology of leptospirosis in Iowa swine was examined on the basis of serologic results and herd data from 55 herds in the National Animal Health Monitoring System (NAHMS) program and culture results and histories from 578 cases of reproductive failure submitted to the Iowa Veterinary Diagnostic Laboratory during a 3-year period. Thirty-eight percent of sera from NAHMS herds contained antibodies against 1 or more of 12 leptospira antigens. Leptospires were isolated from 9 (1.6%) of 578 cases of reproductive failure. Seven (78%) of the isolates were identified as Leptospira interrogans serovar kennewicki and 2 (22%) as serovar grippotyphosa. In 7 herds from which leptospires were isolated, attack rates ranged from 1% to 84%. Clinical leptospirosis, characterized by reproductive failure and confirmed by isolation of leptospires, was sporadic. No significant differences in farrowing averages and reproductive problems were observed between vaccinated and nonvaccinated NAHMS herds or between herds with higher (43-63%) or lower (14-40%) percentages of animals that were serologically positive against serovar bratislava.

Abortion, Veterinary↗

Sporadic cases and an outbreak of leptospirosis probably associated with recreational activities in rivers in the northern part of Okinawa Main Island.

In the summer of 2003, sporadic cases and an outbreak of human leptospirosis probably related to recreation in rivers occurred in the northern part of Okinawa Main Island. Sixteen of 22 suspected cases were definitely diagnosed as leptospirosis by serological test or isolation. The infective leptospiral serovar in 14 cases was presumed to be Hebdomadis. Transmission was thought to occur by exposure to river water that was contaminated by the urine of infected animals. The findings indicate that recreation in rivers in this area is a significant risk factor for infection with leptospires.

Adolescent↗

Prevention and treatment of leptospirosis.

Leptospirosis is a common zoonosis acquired by exposure to body fluids, to tissues of infected animals, or to contaminated soil or fresh water. Large outbreaks of leptospirosis have occurred following excess rainfall or in tourists following exposure to fresh water during adventure activities. Prevention may be achieved through the use of protective clothing or by changes in animal husbandry. Immunization is not widely used in human populations but has a role in agriculture. Oral administration of doxycycline (Periostat , CollaGenex) once-weekly can give short-term protection in high-risk environments. The disease in humans has a broad range of symptoms. Treatment may be symptomatic or may require administration of antibiotics, such as doxycycline or penicillin.

Animals↗

[Involvement of the nervous system in leptospirosis. II. Cerebrospinal fluid evaluation].

From January 1st up to September 30th 1990, 77 patients with leptospirosis diagnosis confirmed by laboratory studies were admitted at the Infectious and Parasitic Disease Service of the Hospital das Clínicas of the Universidade Federal de Pernambuco. The neurological exam was abnormal in 70 of the patients (90.91%). The cerebrospinal fluid (CSF) examination was carried out on 67 (87%) of the patients; it was abnormal in 64 (95.52%) yielding hypercytosis in the majority of cases. There was no difference in the CSF behavior in report to the several neurological forms nor in relation to the icteric or non-icteric forms of the disease. Significant were the results for the microscopic agglutination tests for leptospirosis in the CSF.

Central Nervous System Diseases↗

[Immunodiagnosis of human leptospirosis by ELISA-IgM, employing different antigenic preparations from prevalent serovars of Leptospira interrogans].

A comparative study among different serovars of Leptospira interrogans was performed in order to prepare antigens to detect IgM antibodies by ELISA in early and late phase of human leptospirosis. Ten serovars were chosen among the most prevalent detected by microscopic seroagglutination (SAM) in São Paulo city. Using ELISA-IgM five of them showed better results (canicola, hebdomadis, icterohaemorrhagiae, cynopteri and brasiliensis). These ones were also studied in a pool. The non-treated antigens showed higher reactivity than the Triton X-100 (4%/50 degrees C/4h). ELISA-IgM using individually or pool of non-treated antigens proved to be reliable with high sensitivity and should be used for an earlier diagnosis of leptospirosis, as a trial test. Faster diagnostic elucidation can be useful to detect epidemic situations, so, allowing epidemiological surveillance interventions.

Antigens, Bacterial↗

[An immunoenzyme test (ELISA) for the detection of circulating class-IgA antibodies in human leptospirosis].

An enzyme-linked immunosorbent assay ELISA was evaluated for the detection of IgA antibodies in the human leptospirosis. The assay proved to be sensitive and specific when compared with the ELISA-IgM, in the examined serum samples. The results found suggest that IgA antibodies became positive later in leptospirosis, and will can be an evolutive indicator in the development of the disease.

Adolescent↗

Massive ocular hemorrhage resulting in blindness in a patient with the sickle cell trait who developed leptospirosis. Case report.

This case report describes the findings of a 18 year-old black male from Bahia, a Northeastern state in Brazil, with the sickle cell trait, who developed bilateral hyphema and vitreous hemorrhage with blindness in the course of leptospirosis. The patient started to complain of blurred vision four days after the start of fever and muscular pain and approximately twelve hours after the introduction of penicillin. The severity of the leptospirosis in conjunction with sickle cell trait was considered to be the most likely explanation for this ocular complication.

Adolescent↗

Human leptospirosis: a twenty-nine-year serological study in São Paulo, Brazil.

A retrospective study of 9,335 cases of human leptospirosis in the state of São Paulo, Brazil, diagnosed between 1969 and 1997 showed that the disease is endemic throughout the state. Middle-aged adults, with a range of 20-39 years, were most frequently infected (32.40%). The mean annual incidence was 0.53 per 100,000 population and the disease was more frequent in males (87.0%). Cases occurred mainly in January to April each year. A peak was observed in 1991 and 1996 which rainfall average was 159.9 and 160.3, respectively. These data emphasize the potential public health importance of leptospirosis in the state of São Paulo, Brazil.

Adolescent↗