Bilateral facial palsy and meningitis caused by Borrelia double infection.
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Borrelia↗
Biomedical subjects
M K Viljanen
Publications and source records attributed to M K Viljanen.
Antibodies against whole sonicated Borrelia burgdorferi spirochetes, 41-kilodalton flagellin, and P39 protein in patients with PCR- or culture-proven late Lyme borreliosis.
The sensitivities and specificities of three enzyme-linked immunosorbent assays (ELISAs) for Borrelia burgdorferi antibodies were compared for 41 patients presenting with symptoms compatible with late Lyme borreliosis (LB) and 37 healthy controls. All subjects were living in southwestern Finland, where LB is endemic. Only patients with culture- or PCR-proven disease were enrolled in the study. The antigens of the ELISAs consisted of sonicated spirochetes, 41-kDa flagellin, and recombinant P39 protein of B. burgdorferi. Fifteen patients had strongly or moderately positive results in the serological assay(s), 19 patients had only weakly positive or borderline antibody levels, and the remaining 7 patients were seronegative by ELISA. The sensitivities of the ELISAs were 78.0% with sonicate antigen, 41.5% with 41-kDa flagellin, and 14.6% with P39 protein. The specificities of the tests were 89.2, 86.5, and 94.6%, respectively. The sonicate antigen ELISA seems to be an effective screening method. These results show that antibodies to B. burgdorferi may be present in low levels or even absent in patients with culture- or PCR-proven late LB. Therefore, in addition to serological testing, the use of PCR and cultivation is recommended in the diagnosis of LB.
Adolescent↗
Tick bites, clinical symptoms of Lyme borreliosis, and Borrelia antibody responses in Finnish army recruits training in an endemic region during summer.
Tick bites, associated clinical symptoms, and antibodies against Borrelia burgdorferi were investigated in 77 Finnish army recruits training during summer in an endemic region and 50 control recruits serving outside the habitat of ticks. During a follow-up of 6 months, 26.9% of the study recruits reported tick bites. None gave a history of erythema migrans. Five study recruits and none in the control group had a combination of two or three nonspecific symptoms compatible with Lyme borreliosis. Three of these five study recruits had been bitten by several ticks. Thirteen (16.9%) study recruits and two (4.0%) control recruits had positive antibody levels against B. burgdorferi in their first serum specimens. No significant change in immunoglobulin G antibody levels was seen between the first and second specimens. Immunoglobulin M antibody levels increased in the sera of 13 (11.9%) study recruits and 1 (3.0%) control recruit. We conclude that recruits training in tick habitats are at high risk of tick bites. All recruits starting their military training in endemic areas should be taught to recognize and remove ticks and advised to wear protective clothing.
Adolescent↗
Effects of thermocyclers and primers on the reproducibility of banding patterns in randomly amplified polymorphic DNA analysis.
The effects of thermocyclers and primers on the reproducibility of banding patterns in randomly amplified polymorphic DNA analysis were tested. Purified Bordetella pertussis DNA was analysed with four primers (12-mer), which did not differ from each other in their GC-content. Three different thermocycler models from two manufacturers were tested. Three of the primers produced consistent banding patterns in separate wells of the reaction plates of individual thermocyclers, and in different thermocycler models. In contrast, the fourth primer showed extensive between-instrument variation, and the within-instrument variation with this primer was acceptable only when the most advanced thermocycler model was used. We conclude that only primers that provide consistent banding patterns in different environments should be chosen for interlaboratory use in randomly amplified polymorphic DNA analysis. The primers showing variability in banding patterns can only be used if they produce consistent results in the thermocycler used.
Base Sequence↗
Sensitive and specific polymerase chain reaction assays for detection of Bordetella pertussis in nasopharyngeal specimens.
Polymerase chain reaction (PCR) assays that amplify segments of a repeated gene element and a toxin promoter gene of Bordetella pertussis were compared with a culture established for the diagnosis of pertussis. Of 44 nasopharyngeal (NP) aspirates collected during a pertussis outbreak, repeated gene element PCR showed a positive result in 21 (48%), including all three patients with positive culture results. Results of toxin promoter gene PCR were positive in eight (18%) cases, and the pathogen was not detected in one patient with a positive culture result. A more sensitive nested PCR assay, based on repeated gene element PCR, was then developed. During a second outbreak two different transportation systems were tested in 146 duplicate NP swabs. Transportation of swabs in empty tubes proved to be better than in transport media for PCR. A total of 190 NP specimens from the two outbreaks were tested, and in 56 the results were shown to be positive by PCR, including all 16 cases confirmed as positive by culture. We conclude that the PCR assay is more sensitive than culture in the diagnosis of pertussis; NP swabbing is a simple, practical, and reliable method of collecting clinical specimens for PCR assays and cultures.
Adult↗
Lyme borreliosis associated with complete flaccid paraplegia.
We report the case of a patient with concomitant Lyme borreliosis and acute paraplegia. The paraplegia was complete, flaccid and of upper motor neurone type. The diagnosis of borreliosis was based on the detection of large amounts of IgM and IgG borrelia antibodies in the acute phase serum and on the complete disappearance of IgM antibody during the review period. IgG borrelia antibodies were also detected in the CSF, but leakage of antibodies from the blood to the intrathecal space could not be ruled out. Lymphocytosis and increased total protein concentration in the CSF were signs compatible with neuroborreliosis. Ceftriaxone therapy effected dramatic recovery of the patient. This case suggests that borreliosis should be considered a possible cause of acute flaccid paraplegia.
Acute Disease↗
Subacute multiple-site osteomyelitis caused by Borrelia burgdorferi.
In a pediatric case of severe multiple-site osteomyelitis caused by Borrelia burgdorferi, the presence of spirochetes in a bone lesion was documented both by culture and by the polymerase chain reaction (PCR). Positive PCR results were also obtained with culture fluid yielding spirochetal growth and with acute-phase serum. Although the disease evidently was a late manifestation of Lyme borreliosis, antibodies to B. burgdorferi were low in titer and were restricted to the IgM class. The distribution of osteomyelitic lesions in multiple bones and the positive PCR results obtained with serum argue for hematogenous spread of the spirochetes. Before the specific diagnosis was established, the patient received several potent antimicrobial drugs, without a favorable outcome. In contrast, therapy with ceftriaxone led to a rapid cure that persisted thereafter. We conclude that infection due to B. burgdorferi must be considered a possible cause of subacute pediatric osteomyelitis.
Bone and Bones↗
Antibodies to filamentous hemagglutinin of Bordetella pertussis and protection against whooping cough in schoolchildren.
A pertussis outbreak was studied prospectively in an elementary school with 39 pupils. All had been immunized with at least three doses of Finnish diphtheria-tetanus toxoid-pertussis vaccine. Diagnosis of pertussis was based on culture, polymerase chain reaction results, and EIA serology using filamentous hemagglutinin (FHA), pertussis toxin, and 69-kDa outer membrane protein as antigens. At the first sampling, 21 children had symptoms suggestive of pertussis, and 18 were healthy. Of the latter, 8 remained healthy without any antibiotic treatment and 9 developed clinical pertussis 1-22 days later. One child developed cough later, but this symptom did not meet criteria for pertussis. The mean levels of IgG, IgM, and IgA antibodies to FHA were significantly higher in 8 healthy children than in 9 children who developed pertussis after the first sampling (P < .001, P = .027, and P = .011, respectively). The results show that antibodies to FHA of Bordetella pertussis in immunized schoolchildren correlate with protection against pertussis.
Adhesins, Bacterial↗
Outcomes of Bordetella pertussis infection in different age groups of an immunized population.
Outcomes of Bordetella pertussis infection were studied in 3 age groups (1-3, 4-6, and 7-15 years) during outbreaks in one day care center (n = 29) and in two elementary schools (n = 210). A total of 76 children were confirmed as having B. pertussis infection; 74 were confirmed by polymerase chain reaction (PCR) and 18 by culture. A positive PCR result was less common in children 1-3 years old than in those 4-6 (P = .006). Asymptomatic B. pertussis infection was more common in preschool children than in schoolchildren (P < .001), whereas schoolchildren presented with symptoms compatible with clinical pertussis more often than preschool children (60/210 vs. 3/29, P = .029). In the community of 12,691, the attack rate (laboratory-confirmed patients/100,000) was 317 in children < 4 years, 1838 in children 4-6 years, 2535 in children 7-15 years, and 248 in persons > 15 years. The protection provided by conventional pertussis vaccine is evidently rather short-lived, and booster vaccinations are needed to eradicate pertussis among schoolchildren and adults.
Age Factors↗
Identification of mycobacteria by PCR-based sequence determination of the 32-kilodalton protein gene.
In this study, a part of the nucleotide sequence of the mycobacterial 32-kDa protein gene was determined by PCR-based sequencing. A total of 24 mycobacterial strains, representing 10 species, were studied. Sequences of all tested members of the Mycobacterium tuberculosis complex were identical to each other and to the previously published sequence of M. tuberculosis H37Rv. The sequences of M. avium and M. intracellulare were different from each other. MAIX strains, identified with the Gen-Probe MAIX test, had sequences identical to each other but clearly different from those of M. avium and M. intracellulare. Each of the other mycobacterial species investigated, i.e., M. kansasii, M. gastri, M. gordonae, and M. malmoense, had a unique species-specific sequence. These results demonstrate that there is variation in the nucleotide sequence of the 32-kDa protein gene among different mycobacterial species. Thus, we propose that this gene can be used for PCR-based identification of mycobacteria.
Bacterial Proteins↗
Primers are decisive for sensitivity of PCR.
A sufficient sensitivity of PCR is a prerequisite for its use in the diagnosis of infectious diseases. We have used PCR for detecting gene elements of Borrelia burgdorferi, mycobacteria and Bordetella pertussis. With all these microbe groups, difficulties were encountered in achieving the demanded sensitivity with the primer pairs primarily selected. An extensive testing of various reaction parameters did not improve the sensitivity. Subsequently, we synthesized more primers derived from slightly different positions of the original target sequences. When the original and new primers were tested in possible combinations, some primer pairs reached 100-fold to 1000-fold higher sensitivity than the primary pairs. We conclude that in optimizing the sensitivity of PCR, more emphasis should be put on testing of several primer pairs than on the extensive screening of reaction parameters. Thus far, a trial-and-error approach has to be used, because there is no means to predict the sensitivity properties of a selected primer pair.
Base Sequence↗
Misdiagnosis of Mycobacterium avium-intracellulare by rapid PCR/DNA probe tests.
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DNA Probes↗
Cultivation of Borrelia burgdorferi from the blood and a subcutaneous lesion of a patient with relapsing febrile nodular nonsuppurative panniculitis.
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Adult↗
Detection and identification of mycobacteria by amplification of a segment of the gene coding for the 32-kilodalton protein.
A polymerase chain reaction (PCR) assay for the rapid detection of mycobacterial DNA is described. Oligonucleotide primers, derived from the sequence of a gene coding for the 32-kDa antigen of Mycobacterium tuberculosis, amplified DNA from all 28 species of mycobacteria tested. All nonmycobacterial species tested were negative. An oligonucleotide probe hybridized to the PCR products of the strains belonging to the M. tuberculosis complex. This method could detect as little as 50 fg, as tested with purified M. tuberculosis DNA. By this amplification method, 127 sputum specimens were tested, with 7.9% of the specimens proving to be inhibitory in PCR. The sensitivity of detection by PCR compared with that by culture was 55.9%; when the inhibitory specimens were excluded, the sensitivity was 70.4%. The specificity of PCR combined with hybridization was 100%.
Bacterial Proteins↗
Antibodies to arthritis-associated microbes in inflammatory joint diseases.
IgM, IgG and IgA class antibodies against Yersinia, Salmonella, Campylobacter and Borrelia were determined by enzyme-linked immunosorbent assay (ELISA) in a group of 340 unselected patients with a recent inflammatory joint disease. The control group consisted of 340 and 100 healthy blood donors using Borrelia-ELISA and other ELISAs, respectively. Of all the patients, 27.4% had increased antibody levels against at least one of the microbes tested. The prevalence of positive antibody levels was highest in Yersinia antibodies (17.9%). The corresponding figures for Salmonella, Campylobacter and Borrelia were 7.0, 6.2 and 1.8%, respectively. Patients with entero-arthritis or clinically typical reactive arthritis who had not had gastrointestinal or urogenital symptoms previously had the highest prevalence of the microbial antibodies (67.6 and 40.7%, respectively). These findings indicate that arthritis may often have a reactive etiopathogenesis without recognized gastrointestinal infection, emphasizing the importance of microbial serology in the differential diagnosis.
Antibodies, Bacterial↗
Outbreak of diarrhoea due to Escherichia coli O111:B4 in schoolchildren and adults: association of Vi antigen-like reactivity.
During six days in November, 1987, 611 pupils (age range 7-19 years) and 39 adults (23-57) at a school complex in southern Finland had diarrhoea due to Escherichia coli O111:B4. Diarrhoea developed in 137 other household members during the two weeks after the school outbreak. The source of the organism remains unknown. The outbreak strains, when incubated at 22 degrees C or exposed to ampicillin, lost the lipopolysaccharide O antigen and began to react with antisera against Salmonella typhi Vi antigen. The Vi antigen-like reactivity increased the adherence of the organisms to Hep-2 cells. These results indicate that E coli O111:B4, and possibly other enteropathogenic E coli strains, should be considered in the diagnosis of all diarrhoea cases and not only in infantile diarrhoea. Expression of Vi antigen in E coli may play a part in virulence by enhancing adherence to the intestinal epithelium.
Adolescent↗
Analysis of the relationship between bacterial adherence and extracellular production of mannose, galactose, glucose and ribose in Staphylococcus epidermidis and Staphylococcus hominis.
Gas-liquid chromatography-mass spectrometry was used to analyze the extracellular extracts of 108 coagulase-negative staphylococcal strains for the presence of mannose, galactose, glucose and ribose, in order to determine whether production of these four monosaccharides, regarded as potential staphylococcal slime components, was associated with the adherence capacity of the individuals strains. A total of 90 Staphylococcus epidermidis and 18 Staphylococcus hominis strains were studied. Using the quantitative spectrophotometric assay, 21 Staphylococcus epidermidis strains were classified as strongly adherent, 12 as moderately adherent, 11 as weakly adherent, and 46 as nonadherent. All 18 Staphylococcus hominis strains were nonadherent. Mannose, galactose, glucose and ribose were detected as the main monosaccharide components in the extracellular extracts of all strains examined. Moreover, the mean relative concentrations of these monosaccharides were essentially the same for the different adherence phenotypes within the species Staphylococcus epidermidis. These results showed that there was no causal connection between the adherence of coagulase-negative staphylococci and the extracellular production of any of the four monosaccharides analyzed.
Bacterial Adhesion↗