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PubMed1986

Laboratory diagnosis of herpes simplex virus infections. Principles guiding the development of rapid diagnostic tests.

While the incidence of many bacterial sexually transmitted diseases appears to be decreasing, the complications and frequency of viral sexually transmitted diseases in developed countries has increased over 10-fold in the last decade. Since 1975 genital herpes simplex virus infections have increased at a rate of 12% in the United Kingdom. Significant advances in our understanding of the epidemiology, natural history, and therapy of symptomatic genital herpes has occurred in the last 5 yr. In order to properly utilize this information, however, more widespread availability of laboratory diagnostic testing for herpes simplex virus is needed. The availability of tissue culture isolation facilities for herpes simplex virus has expanded to many community hospitals. More importantly, rapid assays to detect herpes simplex virus using monoclonal and polyclonal antibodies in enzyme-linked immunosorbent assay and immunofluorescent assays and/or detection of herpes simplex virus deoxyribonucleic acid by hybridization methods have also been developed. Recent studies indicate that these assays approach the sensitivity of viral isolation when samples from mucocutaneous lesions are taken. These rapid assays have also allowed clinicians to more rapidly diagnose serious herpes simplex virus infection such as neonatal herpes and to institute antiviral therapy earlier in the course of disease. Although the specificity of these assays in high prevalence populations appears excellent, few studies have evaluated these assays in populations where the prevalence of herpes simplex virus is low. Rapid assays for herpes simplex virus also appear to have decreased sensitivity (less than 60%) in detecting asymptomatic excretion of herpes simplex virus from the lower genital tract.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Viral↗
PubMed1990

Rapid laboratory diagnosis of human cytomegalovirus infection using monoclonal antibody.

The monoclonal antibody E. 13, recognized an early nonstructure antigen (68 KD) of human cytomegalovirus (HCMV), was used for early detection of HCMV in cell cultures. The virus antigen could be detected by the indirect immunofluorescence staining (IF) technique 6 hours after inoculation of HCMV AD169 and NTUH871233 strains into human embryonic lung fibroblast (HELF) cells growing in shell vials prepared by low speed centrifugation (700xg) at 25 degrees C for 1 hour. The intensity of fluorescence reached a peak 14 hours after virus inoculation and remained at the same level for 72 hours. The feasibility of application of this technique for rapid detection of HCMV in the clinical virology laboratory was tried. A total of 201 urine specimens requesting HCMV isolation were inoculated into HELF cells using the shell vial culture method. The virus growth in the shell vial cultures was detected by the indirect IF technique with the monoclonal antibody E.13 at 16 to 18 hours postinoculation. The results obtained were compared with those of the conventional tube culture method. The sensitivity and specificity of the test vs the conventional tube culture method for detection of HCMV in clinical specimens were 93.06% and 99.22%, respectively. The average time of virus isolation by the conventional culture method was 10.9 days, whereas final results with the shell vial culture method were obtained in only one day. The above results indicate that detection of HCMV early antigen in infected cells with monoclonal antibody E.13 is a sensitive and reliable method for rapid diagnosis of HCMV infections.

Antibodies, Monoclonal↗
PubMed1976

[Pathophysiology and laboratory diagnosis of collagen diseases (author's transl)].

Beginning with lesions of the capillary wall, rheumatoid arthritis is morphologically expressed by an aseptic inflammation of the synovia with following necrosis of the cartilage and the underlying bone tissue. Only little is known regarding the pathogenesis but most porbably immune mechansims are involved. There is evidence for the participation of type IV and type III-reactions i.e. sensitized lymphocytes as well as immune complexes. Equally the etiology of RA is totally unknown. Once the inflammatory process has reached some activity, in most cases it shows the character of a selfpertetuation as in other autoimmune diseases. Laboratory examinations include BSR, blood count, measurement of Fe, Cu, RF, complement, streptolysins, staphylolysins, ANA, and in doubtful cases synovial biopsy. Furthermore, effusion fluid of the joint may be examined for RF, ANA, complement, rhagocytes, crystals, protein content etc.--Among the collagen diseases sensu strictori, systemic lupus erythematosus is the most important. The underlying process may be characterized as vasculitis due to immune complexes and local activation of the complement system, the complexes containing native and/or denatured DNA, and antibodies mostly of the IgG class. Laboratory examinations include the demonstration of ANA but should concentrate on the measurement of DNA-antibodies using radioimmunological or at least a sensitive and specific immunofluorescence technique. The value of the various tests is discussed.

Arthritis, Rheumatoid↗
PubMed1999

Laboratory diagnosis of common viral infections of the central nervous system by using a single multiplex PCR screening assay.

A multiplex PCR assay that detects the four commonest causes of viral meningitis and encephalitis in the United Kingdom (herpes simplex virus [HSV] type 1 [HSV-1], HSV type 2 [HSV-2], varicella-zoster virus [VZV], and enteroviruses) was developed, and its sensitivity was compared with those of similar assays described previously for this application. Compared to the previous assays, this single multiplex PCR assay had higher molecular sensitivities for the detection for each of the viruses and improved utility for routine use in a diagnostic laboratory. The assay was used to test a series of 1,683 consecutive cerebrospinal fluid (CSF) samples between June 1997 and March 1998 inclusively. Viral nucleic acid was detected in 138 (8.2%) of the CSF samples, including enteroviruses in 51 samples, HSV-2 in 33 samples, VZV in 28 samples, and HSV-1 in 25 samples. Compared to the accepted relative incidence of viral etiologies, aseptic meningitis due to HSV-2 infection was high, and in adult female patients with symptoms of aseptic meningitis, HSV-2 was the virus most commonly detected in the CSF.

Adult↗
PubMed2005

Evaluation of LIAISON Treponema Screen, a novel recombinant antigen-based chemiluminescence immunoassay for laboratory diagnosis of syphilis.

The purpose of this study was to evaluate the diagnostic performance of LIAISON Treponema Screen (DiaSorin, Saluggia, Italy), a new automated chemiluminescence immunoassay (CLIA), in comparison with that of rapid plasma reagin (RPR) and the following currently used treponemal tests: hemagglutination test (TPHA), immunoenzymatic assay (EIA), and Western blot (WB). First, a retrospective study was performed with a panel of 2,494 blood donor sera, a panel of 131 clinical and serologically characterized syphilitic sera, and 96 samples obtained from subjects with potentially interfering diseases or conditions. A prospective study was also performed by testing 1,800 unselected samples submitted to the Microbiology Laboratory of the St. Orsola Hospital in Bologna, Italy, for routine screening for syphilis. As expected, RPR was the least specific method, especially when potentially cross-reacting sera were tested. On the contrary, all of the treponemal tests proved to be very specific (99.9%) and they performed with the following sensitivities: 100% (WB), 99.2% (CLIA), 95.4% (EIA), and 94.7% (TPHA).

Antigens, Bacterial↗
PubMed1994

Topical thrombin and acquired coagulation factor inhibitors: clinical spectrum and laboratory diagnosis.

Topical bovine thrombin preparations are used extensively in cardiovascular, neurosurgical, and otolaryngologic procedures. Patients who are treated with these topical thrombin preparations may develop antibodies to bovine coagulation factors that may cross-react with the endogenous human clotting proteins. We have identified four patients with acquired factor inhibitors following exposure to topical thrombin at Duke University Medical Center and summarize these cases in addition to 13 patients previously reported in the literature. In most cases, the inhibitor developed following a second (or subsequent) exposure to topical thrombin. The clinical course was extremely variable, ranging from totally asymptomatic to life-threatening hemorrhage. The most consistent laboratory abnormality was a prolonged bovine thrombin clotting time, which corrected, at least partially, when human thrombin was substituted for bovine thrombin. Some of these patients also developed factor V inhibitors with prolonged prothrombin and activated partial thromboplastin times. Although these patients have prolonged clotting times, they should not be considered "autoanticoagulated," since thromboembolic complications can still occur. Therapeutic intervention is largely empirical and depends on the clinical manifestations of the individual patient.

Administration, Topical↗
PubMed2000

Evaluation of LightCycler PCR for implementation of laboratory diagnosis of herpes simplex virus infections.

Five hundred specimens (288 genital, 192 dermal, and 20 ocular) were extracted by technologists, and the DNA was assayed by LightCycler PCR (DNA polymerase and thymidine kinase [TK] gene targets) and by conventional tube and shell vial cell culture. One hundred fifty-eight confirmed (by cell culture and TK target PCR) positive and LightCycler-positive specimens were detected during the first 30 PCR cycles. LightCycler PCR-positive results for cycles 31 to 45 (39 of 67 [58.2%]) required confirmation by another PCR target (TK). LightCycler PCR is more sensitive (n = 197; 23.1%) than cell cultures (n = 150) for the routine laboratory detection of herpes simplex virus infections.

DNA-Directed DNA Polymerase↗
PubMed1977

[Pulmonary mycoses; laboratory diagnosis, antimycotic therapy].

The microscopic, cultural and serological techniques for the diagnosis of European and non-European systemic mycoses involving the respiratory tract are reviewed. The antimicrobial, therapeutic and toxic properties of those antimycotic drugs are discussed, which can be used in the treatment of pulmonary mycoses. Data on biotransformation, kinetics and dosage are reported.

Administration, Topical↗
PubMed2006

Transmission of integrated human herpesvirus 6 through stem cell transplantation: implications for laboratory diagnosis.

We identified a stem cell donor with chromosomally integrated human herpesvirus (HHV)-6 and monitored the recipient for HHV-6 after transplantation. The appearance and subsequent increase in HHV-6 load paralleled engraftment and an increase in white blood cell count. Fluorescent in situ hybridization analysis showed integrated HHV-6 on chromosome band 17p13.3 in the donor and in the recipient after transplantation but not in the recipient before transplantation. The increase in viral load due to the genetic transmission of integrated HHV-6 could have been misinterpreted as substantial active infection and, thus, led to the administration of toxic antiviral therapy. We suggest that the confounding influence of integration be considered in laboratory investigations associating HHV-6 with disease.

Adult↗
PubMed1987

Secondary tuberculosis of the skin. Clinical features and problems in laboratory diagnosis.

One hundred patients with secondary skin tuberculosis--59 with lupus vulgaris (LV), 27 with scrofuloderma (SD), and 14 with tuberculosis verrucosa cutis (TVC)-were included in this study. The buttocks and lower limbs were seen to be important sites of involvement in LV, besides the occurrence over the face. An active focus of tuberculosis was present in 18, a past history of pulmonary tuberculosis in 8, and intrafamilial tuberculous infections in 21. Histopathology and culture for Mycobacterium tuberculosis were done in all the cases. Guinea pig inoculation was done in 11. The poor results of these investigations have been highlighted and discussed with reference to studies done in the past by other workers. The need for improvement in laboratory techniques is suggested.

Adolescent↗
PubMed1994

[Improving the laboratory diagnosis of rabies and titration of rabies viruses].

For primary isolation and titration of street strains of the rabies virus from brains of suspected animals, an assay prepared on the cell culture BHK-21/C 13 (rabies infection test - RTCIT) was used. The above assay proved to be reliable and its sensitivity proved to be comparable to the standard mouse inoculation test. Through this test, the results were obtained within 24 to 48 hrs on Lab-Tek tissue culture chamber/slides. It was found out that DEAE-dextran added to the cell culture only slightly increased the invasiveness of the virus in the samples tested. The method described herein is able to substitute the mouse inoculation test (MIT). In our laboratory, 20 vaccination strains of the rabies virus Vnukovo-32/107 and 25 street strains of the rabies virus (delivered from the field) - original fox brain suspensions. And 10 brain suspensions were negative when tested in laboratory conditions (by PMIF, RTCIT as well as by MIT methods).

Animals↗
PubMed1989

Evaluation of the GO Slide, a growth transport system for the laboratory diagnosis of gonorrhoea.

A growth transport system, the GO Slide (Roche Diagnostica), for the isolation of Neisseria gonorrhoeae from clinical specimens was evaluated. This system was compared with culture on chocolate agar (CA) and modified Thayer-Martin (MTM) agar, and Gram-stained smear for the diagnosis of gonorrhoea. Of 94 urethral specimens from symptomatic males, 77.7% were positive on MTM, 80.9% were positive on CA and Gram stain, and 81.9% were positive on GO Slide. Of the 77 isolates, 26.0% were penicillinase producers, 12.3% belonged to WI serogroup and 87.7% belonged to WII/III serogroup. The ease of transportation and the easy examination of growth on agar surfaces make the GO Slide an attractive system. It would be particularly useful for general practitioners without culture facilities to transport specimens to the laboratory for the isolation of N. gonorrhoeae.

Culture Media↗
PubMedDate not supplied

Laboratory diagnosis of acute myocardial injury.

Considerable effort by clinicians and scientists focuses on the utility of biomarkers to prevent, diagnose and manage adverse cardiac events as well as provide information on a specific patient's underlying pathology. Although troponins I and T (TnI and TnT) are cardiac specific markers that yield diagnostic and prognostic value in patients with myocardial injury, troponins cannot be utilized in all clinical settings. Troponins have limited utility for the diagnosis of early ischemia and preoperative myocardial infarction. Troponin T also lacks specificity in patients with renal failure. New markers, such as ischemia modified albumin (IMA) and CD40 ligand, and new technologies, such as proteomics, are under investigation to advance our knowledge of heart disease.

Albumins↗
PubMedDate not supplied

Tuberculosis: pathogenesis and laboratory diagnosis.

Emerging molecular biologic technologies hold the promise of more rapid diagnosis of tuberculosis and more definitive epidemiologic linkages of cases of TB. In addition to reviewing the pathogenesis of tuberculosis, this chapter covers a variety of methods for the rapid detection of the disease, including the acid-fast smear, conventional culture, the BACTEC system, immunodiagnostic methods, and DNA-based techniques.

Clinical Laboratory Techniques↗
PubMed1995

Laboratory diagnosis of HCMV-related disease in renal transplant patients - pp65 antigen detection versus nested PCR.

BACKGROUND: Sixty-five renal transplant (Tx) recipients were monitored for signs and symptoms of human cytomegalovirus (HCMV) infection. OBJECTIVES: Different diagnostic markers were evaluated for early and correct diagnosis of HCMV disease. STUDY DESIGN: Blood and urine samples were obtained in weekly intervals and the following markers were determined: (1) IgG and IgM antibodies in serum using immunofluorescence and ELISA tests; (2) viral shedding in urine by rapid centrifugation culture (RCC); (3) viral antigen (pp65) in peripheral blood leukocytes (PBL) by immunofluorescence and (4) viral DNA in PBL by nested PCR (NPCR). RESULTS: Twenty-two patients remained free of HCMV infection, 18 patients developed clinical symptoms of HCMV disease, and 25 patients remained asymptomatic in spite of laboratory signs of HCMV infection. For the early detection of HCMV disease, the highest sensitivity was achieved using NPCR (100%) and pp65 antigen detection (94%). RCC and IgM serology were less sensitive (62% and 40% respectively). The differences of sensitivity were significant. Clinical specificity was 47% for NPCR, 79% for pp65 antigen detection, 66% for RCC, and 68% for IgM serology. CONCLUSION: In contrast to NPCR, pp65 antigen detection was closely correlated with the appearance of clinical disease and proved to be a useful marker in the monitoring of antiviral therapy.

Journal Article↗
PubMed2003

Modern laboratory diagnosis of tuberculosis.

One-third of the global population is believed to be infected with bacteria of the Mycobacterium tuberculosis complex, the causative agent of tuberculosis. More than 8 million new cases of tuberculosis occur annually leading to 2 million deaths. Mortality is particularly high in those coinfected with HIV and where the bacteria are multiple-drug-resistant strains--ie, strains resistant to at least isoniazid and rifampicin. Early diagnosis of tuberculosis and drug resistance improves survival and by identifying infectious cases promotes contact tracing, implementation of institutional cross-infection procedures, and other public-health actions. This review addresses significant advances made in the diagnosis of infection, clinical disease, and drug resistance over the past decade. It proposes operational criteria for a modern diagnostic service in the UK (as a model of a low-incidence country) and explores some of the economic issues surrounding the use of these techniques.

Antitubercular Agents↗