Shell Vial Cell Culture Assay Research Articles

Comparison of the “ProDect BCS RV CHIP” assay with the combination of shell vial cell culture and immunofluorescence antibody test for the detection of respiratory viruses

In the present study, a multiplex reverse transcriptase polymerase chain reaction combined with a chip hybridization assay (ProDect BCS RV CHIP) was evaluated as an alternative to the combination of immunofluorescent antibody test and shell vial cell culture considered as gold standard for the detection of respiratory viruses. Among 100 specimens, 40 were positive using the combination of immunofluorescent antibody test and shell vial cell culture assay in which 9 of them were infected by two different viruses (27 parainfluenza virus type 3, 10 adenovirus, 9 respiratory syncytial virus, 2 influenza type B, and 1 influenza type A). ProDect BCS RV CHIP detected only 10 positive specimens in which one of them was infected by two different viruses (5 respiratory syncytial virus, 3 parainfluenza virus type 3, 2 adenovirus, and 1 influenza virus type B). The sensitivity, specificity, PPV, NPV, and diagnostic accuracy of ProDect BCS RV CHIP were 25.0%, 100%, 100%, 66.6%, and 70.0%, respectively, compared to the combination of shell vial cell culture and immunofluorescent antibody test. As a result, the specificity of ProDect BCS RV CHIP is high, however, the sensitivity (25%) of the assay is not sufficient for routine laboratory use.

Identification of Polyoma BK Virus in Kidney Transplant Recipients by Shell Vial Cell Culture Assay and Urine Cytology

Urine cytology often is used to identify BK virus in kidney transplant recipients in the cytology laboratory. To assess the usefulness of the shell vial cell culture assay to identify BK virus, urine samples from 42 kidney transplant recipients were tested by the urine cytology and shell vial cell culture assays. The shell vial cell culture assay is just as sensitive and specific as urine cytology for the identification of BK virus in kidney transplant recipients.

Identification of Polyoma BK Virus in Kidney Transplant Recipients by Shell Vial Cell Culture Assay and Urine Cytology

Urine cytology often is used to identify BK virus in kidney transplant recipients in the cytology laboratory. To assess the usefulness of the shell vial cell culture assay to identify BK virus, urine samples from 42 kidney transplant recipients were tested by the urine cytology and shell vial cell culture assays. The shell vial cell culture assay is just as sensitive and specific as urine cytology for the identification of BK virus in kidney transplant recipients.

Use of Shell-Vial Cell Culture Assay for Isolation of Bacteria from Clinical Specimens: 13 Years of Experience

The shell-vial culture assay is performed routinely in our laboratory. Recently we revisited our experience of using the shell-vial culture assay for the isolation of microorganisms from various clinical samples. Over a 13-year period, we have isolated 580 bacterial strains (5%) from 11,083 clinical samples tested. Over the same period, 285 isolates of rickettsiae, bartonellae, or Coxiella burnetii were cultured from a total of 7,102 samples tested. These isolates include 55 Rickettsia sp. isolates, 95 Coxiella burnetii isolates, and 135 Bartonella sp. isolates. Based on our experience with the growth of fastidious microorganisms, we have used a centrifugation shell-vial technique called JNSP, for "je ne sais pas" ("I don't know [what I am growing]") for the isolation of other microorganisms. A total of 173 isolates were cultured from the 3,861 clinical samples tested using the JNSP method. Of these, 40 isolates had not been grown before on usual axenic medium. These include 2 Staphylococcus aureus isolates, 7 isolates of Streptococcus sp. and related genera, 6 Mycobacterium sp. isolates, 1 Nocardia asteroides isolate, 1 Actinomyces sp. isolate, 1 Brucella melitensis isolate, 2 Francisella tularensis isolates, 1 Mycoplasma pneumoniae isolate, and 1 Legionella pneumophila isolate. Using this protocol, we have also cultured intracellular bacteria such as Chlamydia trachomatis and we have performed the first culture and establishment of Trophyrema whipplei. Applied in our laboratory, the shell-vial culture generally exhibits a low rate of success. However, in some cases, this technique allowed microbial diagnosis when classical agar procedure and PCR were negative.

Seven years' experience of isolation of Rickettsia spp. from clinical specimens using the shell vial cell culture assay.

Annals of the New York Academy of SciencesVolume 990, Issue 1 p. 371-374 Seven Years' Experience of Isolation of Rickettsia spp. from Clinical Specimens Using the Shell Vial Cell Culture Assay G. VESTRIS, G. VESTRIS Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorJ. M. ROLAIN, J. M. ROLAIN Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorP. E. FOURNIER, P. E. FOURNIER Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorM. L. BIRG, M. L. BIRG Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorM. ENEA, M. ENEA Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorJ. Y. PATRICE, J. Y. PATRICE Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorD. RAOULT, Corresponding Author D. RAOULT Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceAddress for correspondence: Didier Raoult, Unité des Rickettsies, Faculté de Médecine, Université de la Méditerranée, 27, Boulevard Jean Moulin, 13385 Marseille Cedex 5, France. Voice: 33-04-91-32-43-75; fax: 33-04-91-38-77-72; Didier.Raoult@medecine.univ-mrs.fr.Search for more papers by this author G. VESTRIS, G. VESTRIS Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorJ. M. ROLAIN, J. M. ROLAIN Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorP. E. FOURNIER, P. E. FOURNIER Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorM. L. BIRG, M. L. BIRG Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorM. ENEA, M. ENEA Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorJ. Y. PATRICE, J. Y. PATRICE Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceSearch for more papers by this authorD. RAOULT, Corresponding Author D. RAOULT Unité des Rickettsies CNRS UMR-A 6020, IFR 48, Faculté de Médecine, Université de la Méditerranée, 13385 Marseille Cedex 05, FranceAddress for correspondence: Didier Raoult, Unité des Rickettsies, Faculté de Médecine, Université de la Méditerranée, 27, Boulevard Jean Moulin, 13385 Marseille Cedex 5, France. Voice: 33-04-91-32-43-75; fax: 33-04-91-38-77-72; Didier.Raoult@medecine.univ-mrs.fr.Search for more papers by this author First published: 18 July 2008 https://doi.org/10.1111/j.1749-6632.2003.tb07394.xCitations: 24Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume990, Issue1RICKETTSIOLOGY: Present and Future DirectionsJune 2003Pages 371-374 RelatedInformation

Evaluation of the COBAS AMPLICOR CMV MONITOR test for detection of viral DNA in specimens taken from patients after liver transplantation.

Detection of cytomegalovirus (CMV) DNA in blood by PCR is a sensitive method for the detection of infection in patients posttransplantation. The test, however, has low specificity for the identification of overt CMV disease. Quantitative CMV PCR has been shown to overcome this shortcoming. The COBAS AMPLICOR CMV MONITOR test was evaluated by using consecutive serum and peripheral blood mononuclear cell (PBMN) samples from liver transplant patients. Twenty-five patients had CMV viremia (by shell vial cell culture assay) and/or tissue-invasive disease (by biopsy); 20 had no active infection. A total of 262 serum and 62 PBMN specimens were tested. Of 159 serum specimens from patients with overt CMV infection, the COBAS assay detected CMV DNA in 21 patients (sensitivity, 84%). Only 1 of 103 samples from patients with no evidence of active infection had detectable CMV DNA (341 copies/ml). By comparison of 62 matching serum and PBMN samples by the same assay, 12 PBMN samples were exclusively positive, whereas only 2 serum samples were exclusively positive (P < 0.05). At the time of clinical CMV infection, viral copy numbers were higher in PBMNs than serum from four of five patients. The COBAS AMPLICOR CMV MONITOR test is a sensitive and specific test for the quantitative detection of CMV DNA in blood. Clinical applications of the assay will require further validation with samples from a larger population of transplant patients.

Survey of urine from transplant recipients for polyomaviruses JC and BK using the polymerase chain reaction

Using polymerase chain reaction (PCR), we examined 108 urine specimens from 39 post transplant patients for polyomaviruses JC (JCV) and BK (BKV). Urine sediments were collected and subjected to 30 cycles of amplification. PCR products were resolved by agarose gel electrophoresis, transferred to nylon membranes by Southern blot, and hybridized with radiolabelled probes. Polyomavirus DNA was found in urine specimens from 17 out of 39 patients (44%). Both viruses were detected in specimens from nine patients, JCV alone in five, and BKV alone in three. In comparison, polyomavirus was detected in only five of 22 PCR positive specimens by shell vial cell culture assay. Our results show a high prevalence of polyomavirus shedding after transplantation and suggest a higher rate of JC viruria than previously reported.

Rapid detection of polyomavirus BK by a shell vial cell culture assay

Polyomavirus BK (BKV) causes asymptomatic latent infection in the human host that is reactivated during periods of immune suppression. Detection by conventional tube cell culture is difficult and time consuming because BKV exhibits slow growth with late (14 to 28 days) and subtle cytopathic effects. We developed a shell vial cell culture assay (SVA) using a cross-reactive monoclonal antibody to the T antigen of simian virus 40 to detect BKV rapidly by indirect immunofluorescence. Nuclear fluorescence was seen in BKV-infected cells as early as 16 h postinoculation; 6 to 28 times more foci were present at 36 h postinoculation. Human embryonic kidney cells infected with BKV produced 7 to 42 times more fluorescent foci than MRC-5 or rhabdomyosarcoma cells did. Centrifugation enhanced the infectivity of BKV in the SVA. To define the clinical utility of SVA, urine specimens from organ transplant patients were tested. Of 27 patients, 4 (15%) were found to be positive by SVA. SVA offers a simple and rapid method for detection of BKV that can be of use in clinical studies of this virus.

Rapid Shell Vial Culture and Tissue Histology Compared With Serology for the Rapid Diagnosis of Cytomegalovirus Infection in Liver Transplantation

Sixty-six patients who had undergone 78 liver transplantations, with no detectable cytomegalovirus (CMV) infection before transplantation, were studied to assess the value of CMV serology compared with the rapid detection of the virus in shell vial cell cultures or histology for the diagnosis of (1) the first evidence of infection, (2) symptomatic disease, and (3) asymptomatic infection. Of 28 evaluable patients, shell vial cell culture assay and histologic findings provided the first evidence of CMV infection earlier in 22 (79%) or at the same time as the serologic response in 5 (18%). Serologic results yielded the first indication of CMV infection in only 1 of these 28 patients (3%). Of 17 evaluable transplantations in 15 patients in whom symptomatic invasive disease developed, shell vial culture or histology provided a laboratory diagnosis of CMV infection earlier or at the same time as serologic responses in 16 (94%). Similarly, shell vial culture or histology provided evidence of CMV infection in 10 of 11 patients (91%) who had asymptomatic CMV infection and remained asymptomatic. Urine surveillance cultures yielded the first evidence of CMV infection in 14 of 19 patients who initially had an asymptomatic infection, of which 6 became symptomatic. Culturing of the blood with use of the rapid shell vial technique showed that viremia preceded CMV organ involvement in 7 of 10 patients. We recommend culture by the shell vial assay as the most rapid and sensitive method of determining CMV infection in liver transplant patients.

Detection of IgM antibody to cytomegalovirus and rapid diagnosis of this virus infection by the shell vial assay

Multiple serum specimens from 10 patients with known cytomegalovirus (CMV) infections and 498 sera consecutively submitted to the laboratory for the diagnosis of CMV infection were tested for anti-CMV IgM after treatment with goat antihuman IgG and QAE-Sephadex A50 column chromatography. Specimens from all 10 patients were positive for IgM after treatment with anti-IgG, but only 8 after the column procedure. Anti-CMV IgM was detected in 23 of 498 (4.6%) specimens pretreated with anti-IgG but in only 12 (2.4%) of these samples after the sera was passed through QAE-Sephadex A50 columns. Anti-CMV IgM was detected exclusively in 2 sera after QAE-Sephadex A50 column treatment (sensitivity, 83%) and in 13 specimens pretreated with anti-IgG sera (specificity, 97%). Serology (IgG and IgM) was compared with the shell vial cell culture assay and histology for providing the first evidence of CMV infection in 28 liver transplant patients. CMV infection was detected initially by the rapid shell vial assay (24) or histology (3) in 27 (96%) of these patients. Detection of anti-CMV IgM in these patients had little value for rapid diagnosis of these infections, and suggests that serology should be recommended mainly for the diagnosis of primary CMV infections in localities in which the rapid shell vial assay is not available.