Detection of multiple reactive protein species by immunoblotting after recombinant outer surface protein A lyme disease vaccination.

Laboratory confirmation of the diagnosis of Lyme disease is based on the detection of an immune response to Borrelia burgdorferi. The serodiagnosis of B. burgdorferi infection is complex and may be further confounded by the immune response to the recombinant outer surface protein A (OspA) Lyme disease vaccine. To describe how the serological response to the recombinant OspA Lyme disease vaccine affects testing for antibody to B. burgdorferi, 240 specimens from 80 study subjects were obtained at defined intervals after recombinant OspA Lyme disease vaccination. Samples were tested by indirect enzyme-linked immunosorbent assay (ELISA), antibody capture enzyme immunoassay (EIA), and Western blotting (WB). After recombinant OspA Lyme disease vaccination, ELISA for 98% of the study subjects revealed reactivity. WB with use of OspA-containing B. burgdorferi strains as sources of antigens demonstrated multiple bands. Results of testing with a US Food and Drug Administration-approved WB kit showed homogeneous reactivity in the molecular weight region >30 kDa. Testing with OspA-free strains completely eliminated all vaccine-associated reactivity by both antibody capture EIA and WB.

The long-term clinical outcomes of Lyme disease. A population-based retrospective cohort study.

OBJECTIVE: To ascertain the prevalence of and risk factors for long-term sequelae from acute Lyme disease. DESIGN: Population-based, retrospective cohort study. SETTING: A coastal region endemic for Lyme disease. PARTICIPANTS: Patients with a history of Lyme disease who were previously treated with antibiotics were compared with randomly selected controls. MEASUREMENTS: A standardized physical examination, health status measure (Short Form 36), psychometric test battery, and serologic analysis. RESULTS: Compared with the control group (n = 43), the Lyme group (n = 38; mean duration from disease onset to study evaluation, 6.2 years) had more arthralgias (61% compared with 16%; P < 0.0001); distal paresthesias (16% compared with 2%; P = 0.03); concentration difficulties (16% compared with 2%; P = 0.03); and fatigue (26% compared with 9%; P = 0.04), and they had poorer global health status scores (P = 0.04). The Lyme group also had more abnormal joints (P = 0.02) and more verbal memory deficits (P = 0.01) than did the control group. Overall, 13 patients (34%; 95% CI, 19% to 49%) had long-term sequelae from Lyme disease (arthritis or recurrent arthralgias [n = 6], neurocognitive impairment [n = 4], and neuropathy or myelopathy [n = 3]). Compared with controls, patients who had long-term sequelae had higher IgG antibody titers to the spirochete (P = 0.03) and received treatment later (34.5 months compared with 2.7 months; P < 0.0001). CONCLUSIONS: Persons with a history of Lyme disease have more musculoskeletal impairment and a higher prevalence of verbal memory impairment when compared with those without a history of Lyme disease. Our findings suggest that disseminated Lyme disease may be associated with long-term morbidity.

Evaluation of the intrathecal antibody response to Borrelia burgdorferi as a diagnostic test for Lyme neuroborreliosis.

The intrathecal antibody response to Borrelia burgdorferi was evaluated in American and West German patients with Lyme neuroborreliosis. By an antibody capture enzyme immunoassay, 12 (92%) of 13 patients from the USA with Lyme meningitis were found to have intrathecal antibody production to B. burgdorferi, usually of multiple isotypes, most commonly IgA. Of 12 patients with putative late central nervous system manifestations of Lyme disease, 5 (42%) had local production of IgG or IgA spirochetal antibody, but cerebrospinal fluid (CSF) abnormalities could not be demonstrated in 6 patients with late peripheral nervous system manifestations of the disorder. Compared with American patients, 30 European patients with neuroborreliosis had significantly higher CSF:serum ratios of specific antibody both early and late in the illness. Intrathecal antibody determinations are the most specific diagnostic test currently available for Lyme neuroborreliosis, but local antibody production in CSF is an inconsistent finding in American patients with late neurologic manifestations of the disorder.

Rapid emergence of a focal epidemic of Lyme disease in coastal Massachusetts.

We describe a focal epidemic of Lyme disease, which spread from a nature preserve and affected an adjacent community of permanent residents in coastal Massachusetts. The attack rate from 1980 through 1987 was 35 percent among 190 residents living within 5 km of the nature preserve and was greatest (66 percent) among those living closest to the preserve. The risk of infection bore little relation to sex or age. Late Lyme disease, which clustered near the preserve, occurred mainly in residents infected early in the epidemic who did not have a history of erythema migrans and did not receive antibiotic therapy. All the residents with serologic evidence of infection had early or late clinical manifestations of Lyme disease, or both, during the period of study. The seasonal risk of infection was bimodal--greatest in June, with a secondary peak in October--and corresponded to periods of increased transmission. In the nature preserve, the density of the vector tick, Ixodes dammini, exceeded that in other New England sites. The zoonosis rapidly became endemic, and the severity of its impact correlated with the abundance of deer. This epidemic of Lyme disease demonstrated that outbreaks can be focal and can spread rapidly within a community of permanent residents.

Serodiagnosis of early Lyme disease: analysis of IgM and IgG antibody responses by using an antibody-capture enzyme immunoassay.

We used an antibody-capture enzyme immunoassay (EIA) to evaluate the early antibody responses to Borrelia burgdorferi in paired sera from 30 patients with erythema chronicum migrans. During acute disease, 20 (67%) patients had elevated specific IgM responses, and by convalescence (one to four weeks after treatment), 28 (93%) patients had increased IgM or IgG responses. In acute specimens, elevated IgM responses correlated with disseminated infection; however, by convalescence, most patients with either localized or disseminated disease had positive tests. Among 133 control subjects, IgM cross-reactivity was observed in 4 of 37 patients with either Epstein-Barr virus or rickettsial infections, and false-positive IgG tests were seen in 8 of 28 patients with syphilis. With antibody-capture EIA, the diagnosis of Lyme disease can be confirmed in the majority of acutely ill patients and in almost all patients by convalescence.

Lyme borreliosis in the Soviet Union: a cooperative US-USSR report.

We identified 90 patients with tick-borne erythema migrans in the Union of Soviet Socialist Republics (USSR) in areas from the western Baltic Republics to the Maritime Territory on the Pacific Ocean. Symptoms associated with the erythema included fever, malaise and fatigue, headache, myalgias, arthralgias, or regional lymphadenopathy. Within two weeks to four months, 58 (64%) of the patients developed neurological abnormalities, particularly radicular pain, cranial neuritis, or lymphocytic meningitis, and four (4%) patients developed monoarticular or oligoarticular arthritis. We tested the sera from 35 Soviet patients by using an isolate from the United States. The serological data showed elevated IgM and/or IgG antibody titers to Borrelia burgdorferi in 2 of 10 patients with erythema migrans, 15 of 21 with neurological abnormalities, and 2 of 4 with arthritis. Our observations suggest that Lyme borreliosis occurs in diverse areas of the USSR.

Eastern equine encephalitis in Massachusetts: a report of 16 cases, 1970-1984.

We studied the case records of 16 patients with eastern equine encephalitis (EEE) in Massachusetts from 1970 to 1984 and compared their presentations, courses, and outcomes with the data available from previous epidemics. In recent years, there has been a greater frequency of EEE in adults, whereas in the past it was considered a disease of children. Also, prognosis for a good functional recovery seems to be correlated with age over 40 years, a long prodromal course (5 to 7 days) of constitutional symptoms, and the absence of coma. Previous reports did not mention these significant correlations. We also stress the positive and negative diagnostic correlations, in order to distinguish between EEE and herpes simplex encephalitis.

Seroprevalence of human immunodeficiency virus among childbearing women. Estimation by testing samples of blood from newborns.

Attempts to predict the course of the epidemic of acquired immunodeficiency syndrome (AIDS) have been hampered by the lack of an objective, practical way to estimate the prevalence of infection with the human immunodeficiency virus (HIV) in the general population. Testing for the prevalence of HIV infection in women should be a sensitive means to track the epidemic and to study the potential for perinatal transmission. Antibodies in maternal blood are contained in neonatal blood specimens routinely collected on absorbent paper for other purposes, such as screening for phenylketonuria; we therefore tested for HIV antibody in these specimens. Analysis of batches of individually blinded specimens from selected hospitals protected the anonymity of the mothers and babies and was cost efficient. Using the newborn's blood as an indicator of the mother's serologic status, we concluded that 1 of every 476 women (2.1 per 1000) giving birth in Massachusetts was positive for HIV antibody by immunofluorescence assay or enzyme-linked immunosorbent assay, both confirmed by immunoblot (Western blot) testing. The prevalence of HIV infection varied according to the type and location of the maternity hospitals; rates of seropositivity were highest in inner-city hospitals (8.0 per 1000), lower in mixed urban and suburban hospitals (2.5 per 1000), and lowest in suburban and rural hospitals (0.9 per 1000). This method is useful for collecting data needed to plan and evaluate prevention strategies and to predict the health care resources that will be needed to care for women and children who contract AIDS. Because other states have newborn screening programs similar to the Massachusetts program, this approach can be used for national surveillance of AIDS in women.

Use of cytomegalovirus immune globulin to prevent cytomegalovirus disease in renal-transplant recipients.

We undertook a prospective randomized trial to examine whether an intravenous cytomegalovirus (CMV) immune globulin would prevent primary CMV disease in renal-transplant recipients. Fifty-nine CMV-seronegative patients who received kidneys from donors who had antibodies against CMV were assigned to receive either intravenous CMV immune globulin or no treatment. The immune globulin was administered in multiple doses over the first four months after transplantation. The incidence of virologically confirmed CMV-associated syndromes was reduced from 60 percent in controls to 21 percent in recipients of CMV immune globulin (P less than 0.01). Fungal or parasitic superinfections were not seen in globulin recipients but occurred in 20 percent of controls (P = 0.05). Only 4 percent of globulin recipients had marked leukopenia (reflecting serious CMV disease), as compared with 37 percent of the controls (P less than 0.01). There was a concomitant but not statistically significant reduction in the incidence of CMV pneumonia (17 percent of controls as compared with 4 percent of globulin recipients). A significant reduction in serious CMV-associated disease was observed even when patients were stratified according to therapy for transplant rejection (P = 0.04). We observed no effect of immune globulin on rates of viral isolation or seroconversion, suggesting that treated patients often harbored the virus but that clinically evident disease was much less likely to develop in them. We conclude that CMV immune globulin provides effective prophylaxis in renal-transplant recipients at risk for primary CMV disease.

Borrelia burgdorferi in joint fluid in chronic Lyme arthritis.

Although indirect evidence suggests that chronic Lyme arthritis is caused by persistent infection with Borrelia burgdorferi, direct visualization has been lacking. We report the demonstration of B. burgdorferi from synovial fluid aspirated from the right knee of a 31-year-old man with Lyme arthritis for more than 1 year. After 6 days, culture medium inoculated with synovial fluid showed one motile and several nonmotile spirochetes. Direct immunofluorescence staining showed reactivity with anti-B. burgdorferi serum. Spirochetes were not seen in subcultured material. The patient's arthritis improved with high-dose intravenous penicillin. Identification of B. burgdorferi from the joint fluid of a patient with long-standing arthritis supports the concept that the arthritis is due to persistent infection.

Specificity of immunoglobulin M and G antibody responses in humans infected with eastern and western equine encephalitis viruses: application to rapid serodiagnosis.

Paired sera from 20 humans with eastern equine encephalitis (EEE) virus infections and from 17 humans with western equine encephalitis (WEE) virus infections, all with previously demonstrated fourfold or greater rises or falls in hemagglutination-inhibiting, complement-fixing, or neutralizing antibody titers, were tested for immunoglobulin M (IgM) and IgG antibodies by an enzyme immunoassay. All individuals with EEE and 14 of 17 individuals with WEE had IgM antibody, some as early as 1 day after onset. Two of the three persons with WEE who did not develop IgM antibody died. IgM antibody declined but persisted for at least 3 months after the onset of illness in one individual each with EEE and WEE. IgG antibody was not detected until the middle of week 2 after onset. The sensitivity of the IgM antibody capture enzyme immunoassay described and the specificity, as shown by the absence of heterologous alphavirus reactivity, indicate that this is the test of choice for the rapid diagnosis of human infections caused by EEE and WEE viruses.