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PubMedDate not supplied

Nonketotic hyperglycinemia (glycine encephalopathy): laboratory diagnosis.

Nonketotic hyperglycinemia (NKH) is an autosomal recessive disorder of glycine metabolism caused by a defect in the glycine cleavage enzyme complex (GCS). GCS is a complex of four proteins encoded on four different chromosomes. In classical neonatal NKH, levels of cerebrospinal fluid (CSF) glycine and CSF/plasma glycine ratio are very high but the CSF results, in particular, may be more difficult to interpret in later-onset, milder, or otherwise atypical NKH. Enzymatic confirmation of NKH requires a liver sample. Delineation of which protein of the complex is defective is necessary to screen for mutations in the appropriate gene. Except for Finnish NKH patients, few recurrent mutations have yet been found, although analysis of the P-protein gene (the site of the defect in the majority of patients) is at an early stage. Prenatal diagnosis by GCS assay in chorionic villus biopsies is not completely reliable and will be replaced by molecular analysis in families where the mutations are known.

Animals↗
PubMed1988

Monoclonal antibodies in the laboratory diagnosis of trachoma.

Various techniques which use monoclonal antibodies to detect Chlamydia trachomatis in clinical specimens are reviewed. An investigation comparing the efficacy of immunofluorescent staining with Giemsa staining in detecting Chlamydia in conjunctival scrapings from cases of active trachoma is presented. Sixty-two eyes of schoolboys with moderate to severe trachoma were studied. Giemsa staining detected chlamydial inclusion bodies in 34 percent of the specimens. Free elementary bodies were detected by fluorescent monoclonal antibody in 21 percent. Eleven percent were positive by both Giemsa and immunofluorescence and 55 percent were positive by either Giemsa and/or immunofluorescence. The addition of fluorescent monoclonal antibody assay to routine Giemsa staining resulted in an increase in the yield of positive specimens by 29 percent.

Antibodies, Monoclonal↗
PubMed1986

Evaluation of laboratory diagnosis of toxoplasmosis by means of an ELISA-triple test. Detection of class specific IgG, IgM and circulating antigen.

The combination of three parameters (IgG, IgM and circulating antigen) in a so-called 'ELISA triple test' was suggested for advantageous diagnosing of human toxoplasmosis. A qualitative assay was used with the following arbitrary assumptions: IgG antibodies reflect an infection, IgM antibodies reflect a recent (primary) infection and circulating antigens reflect an active infection. The three assays were performed simultaneously in one microtiter plate. This approach was tested with 1091 patient sera submitted for routine diagnosis. In comparison with conventional indirect immunofluorescence and complement fixation test it was observed that combinations indicating a recently acquired infection (combinations with IgM and/or circulating antigen) mainly paralleled low or negligible conventional antibody titers. No strict association was seen between particular combinations and certain clinical symptoms suggestive for toxoplasmosis. In conclusion it was stated that the triple test for support of clinical diagnosis has some advantages but that a strong need exists to be able to demonstrate exacerbation or reinfections which are generally not characterised by IgM antibody formation of free circulating antigens but may be the reason for elevated conventional antibody titers.

Antigens, Protozoan↗
PubMed1989

Development of a chromosomal DNA probe for the laboratory diagnosis of aspergillosis.

Chromosomal DNA was extracted from clinical isolates of Aspergillus fumigatus of human and animal origin using the protoplast lysate method. The probe was developed by the nick translation of the chromosomal DNA genome fraction with p32 as the radiolabel. Hybridization of the probe with endonuclease-cleaved DNA of the same species resulted in a pattern of recognition sites specific for the species. The latter was not seen in other species encountered in clinical specimens. Trials were carried out on sputum experimentally inoculated with the fungus where crude DNA was directly extracted, treated with the endonuclease and hybridized with the probe. The efficacy of the probe was as good with the crude as the purified DNA. The specificity of the probe was determined by testing it against single and mixed DNA populations extracted from different species of several fungal and bacterial genera isolated from and/or known to occur in clinical specimens of respiratory infection origin. The sensitivity of the probe was assessed by detecting a DNA concentration in the specimen equivalent to 3 C.F.U.

Animals↗
PubMed1976

The laboratory diagnosis and epidemiology of Mycoplasma pneumoniae in Switzerland.

The analysis of approximately 60,000 serum samples collected from patients with different clinical symptoms between 1965 and 1975 in Switzerland yielded positive reactions indicating a recent infection with Mycoplasma pneumoniae in 2-30% during this period. Among 1,555 patients with clinical symptoms of an acute respiratory infection, 359 patients or 23% had antibody titers indicating a recent infection with M.pneumoniae. On the basis of these results it is concluded that M.pneumoniae is the most frequent cause of acute infections of the respiratory tract.

Antibodies, Bacterial↗
PubMedDate not supplied

Lung abscess caused by Streptococcus pneumoniae type 3. The importance of counterimmunoelectrophoresis in laboratory diagnosis.

We are reporting the case of an 18-month-old girl with a post-pneumonic lung abscess. The counterimmunoelectrophoresis (CIE) of the patient's urine during the bacteremic phase of the disease gave a precipitation line with pneumococcal antiserum. Later in the course of the disease, after the diagnosis of lung abscess had been established, the pus drained from the abscess gave positive reactions with pneumococcal omnivalent serum and type 3 pneumococcal antiserum. At this stage, the pus was sterile and blood cultures were negative. This case demonstrates that the CIE test is a simple procedure which gives a rapid and exact diagnosis up to the serotype level.

Counterimmunoelectrophoresis↗
PubMedDate not supplied

Laboratory diagnosis of congenital human cytomegalovirus infection using polymerase chain reaction and shell vial culture.

Congenital HCMV infection was diagnosed at the 22th week of gestation. The infection was suspected because HCMV IgM was detected in a serum sample obtained from the woman's husband. HCMV infection was detected in the amniotic fluid by polymerase chain reaction, shell vial culture (immunoperoxidase assay) and conventional virus isolation. Serologic testing in paired sera of the woman and in umbilical cord blood for specific IgM and IgA remained negative. As serological data (preconceptional HCMV serostatus) were incomplete, a clear differentiation between primary and secondary infection could not be achieved; consequently, risk quantification could not be determined. Viruria was detected in the offspring during the 1st week post partum. No clinical signs of cytomegalic inclusion disease were diagnosed up to six weeks post partum. Our case report indicates that for pregnancy surveillance, serologic testing for HCMV antibody should also be performed in the spouse.

Adult↗
PubMed1987

Laboratory diagnosis of infections caused by legionellae.

Legionnaires' disease can be diagnosed by detection of antibody to the organism or by direct detection of the bacterium and/or its products using immunological means, culture or a DNA probe. Culture isolation, still the most specific and sensitive test, can be done with sputum samples if proper selective techniques are used. Antibody detection is more suited for epidemiological purposes than individual cases. It has been overused to the exclusion of more specific and rapid methods, such as culture and other means of bacterial detection. Immunofluorescent microscopy remains an important tool in diagnosis, although urinary antigen detection and DNA probes are promising alternative tests. For optimal sensitivity, culture must be performed, regardless of which test is used.

Antibodies, Bacterial↗
PubMed1977

RD cells in the laboratory diagnosis of enteroviruses.

It is shown that, in addition to the already known replication of Coxsackie A viruses, 16 out of 18 different types of ECHO viruses tested as well as all three types of polioviruses can also be grown in RD cells with the development of a pronounced CPE. One of the exceptions, ECHO virus type 16, can be propagated in either HEL or HFDK cells and the other, ECHO virus type 22, in Vero cells. These four permanent or semi-permanent cell cultures thus prove very useful for the isolation of most of the diagnostically important enteroviruses as well as for their serological identification by neutralization in a micro-titer system described in this paper.

Cell Line↗