human pbmcs  (Roche)


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    Structured Review

    Roche human pbmcs
    Viral loads, <t>HBV</t> antigens, and antibodies in coculture of HepG2.2.15 with IL-33-stimulated <t>PBMCs.</t> HepG2.2.15 cells were cocultured alone, treated with IL-33 (1 ng/mL), cocultured with PBMCs, IL-33, and PBMCs or IL-33/PBMCs/anti-IL-21, respectively.
    Human Pbmcs, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmcs/product/Roche
    Average 92 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    human pbmcs - by Bioz Stars, 2020-09
    92/100 stars

    Images

    1) Product Images from "IL-33 Enhances Humoral Immunity Against Chronic HBV Infection Through Activating CD4+CXCR5+ TFH Cells"

    Article Title: IL-33 Enhances Humoral Immunity Against Chronic HBV Infection Through Activating CD4+CXCR5+ TFH Cells

    Journal: Journal of Interferon & Cytokine Research

    doi: 10.1089/jir.2013.0122

    Viral loads, HBV antigens, and antibodies in coculture of HepG2.2.15 with IL-33-stimulated PBMCs. HepG2.2.15 cells were cocultured alone, treated with IL-33 (1 ng/mL), cocultured with PBMCs, IL-33, and PBMCs or IL-33/PBMCs/anti-IL-21, respectively.
    Figure Legend Snippet: Viral loads, HBV antigens, and antibodies in coculture of HepG2.2.15 with IL-33-stimulated PBMCs. HepG2.2.15 cells were cocultured alone, treated with IL-33 (1 ng/mL), cocultured with PBMCs, IL-33, and PBMCs or IL-33/PBMCs/anti-IL-21, respectively.

    Techniques Used:

    Related Articles

    Transfection:

    Article Title: Isolation and Characterization of a Replication-Competent Molecular Clone of an HIV-1 Circulating Recombinant Form (CRF33_01B)
    Article Snippet: .. Replication kinetics in human PBMCs and T cell lines To test the replication kinetics and coreceptor tropism, viral stock was prepared by transfecting 5 µg HIV-1 clone into 6×105 HeLa cells using the FuGENE 6 transfection reagent (Roche Diagnostic GmbH, Penzberg, Germany). ..

    Luciferase:

    Article Title: IL-33 Enhances Humoral Immunity Against Chronic HBV Infection Through Activating CD4+CXCR5+ TFH Cells
    Article Snippet: .. The concentrations of HBV DNA were examined in the serum samples from HBV-Tg mice, and the supernatants of HepG2.2.15 cells cocultured with human PBMCs were quantified by luciferase assays using the luciferase quantization detection kit (Roche Amplicor, Basel, Switzerland), according to the manufacturers' instruction (Jiang and others ). ..

    Isolation:

    Article Title: Use of Inhibitors To Evaluate Coreceptor Usage by Simian and Simian/Human Immunodeficiency Viruses and Human Immunodeficiency Virus Type 2 in Primary Cells
    Article Snippet: .. Human PBMC were isolated from various healthy blood donors by Ficoll-Hypaque separation and stimulated for 3 days with phytohemagglutinin (5 μg/ml) and interleukin-2 (IL-2; 100 U/ml) (a gift from Hofmann-La Roche, Inc., Nutley, N.J.). ..

    Article Title: Heart non-specific effector CD4+ T cells protect from postinflammatory fibrosis and cardiac dysfunction in experimental autoimmune myocarditis
    Article Snippet: .. CD4+ T cell proliferation CD4+ T cells were isolated from mouse erythrocyte-lysed splenocytes/LNs or human PBMCs by MACS or sorted from hearts digested with Liberase (Roche). .. CD4+ T cell subpopulations were isolated using FACS Aria III.

    Article Title: Human T-cell leukemia virus type 2 post-transcriptional control protein p28 is required for viral infectivity and persistence in vivo
    Article Snippet: .. Briefly, freshly isolated human PBMCs were pre-stimulated with 2 μg/ml PHA and 10 U/ml IL-2 (Roche, Indianapolis, IN) for three days. .. 729 producer cells (2 × 103 ) were irradiated (100 Gy) and co-cultured with 104 pre-stimulated PBMCs in the presence of IL-2 in 96-well round bottom plates.

    Article Title: Human T-Cell Leukemia Virus Type 2 Antisense Viral Protein 2 Is Dispensable for In Vitro Immortalization but Functions To Repress Early Virus Replication In Vivo
    Article Snippet: .. Briefly, freshly isolated human PBMCs were prestimulated with 2 μg/ml phytohemagglutinin (PHA) and 10 U/ml IL-2 (Roche, Indianapolis, IN) for 3 days. .. The 729 virus producer cells were irradiated (100 Gy) and cocultured with 104 prestimulated human PBMCs at a ratio of 1:5, respectively, with 10 U/ml IL-2 in 96-well plates.

    Mouse Assay:

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Quantification of vector copy numbers and LM‐PCR Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

    Article Title: IL-33 Enhances Humoral Immunity Against Chronic HBV Infection Through Activating CD4+CXCR5+ TFH Cells
    Article Snippet: .. The concentrations of HBV DNA were examined in the serum samples from HBV-Tg mice, and the supernatants of HepG2.2.15 cells cocultured with human PBMCs were quantified by luciferase assays using the luciferase quantization detection kit (Roche Amplicor, Basel, Switzerland), according to the manufacturers' instruction (Jiang and others ). ..

    Real-time Polymerase Chain Reaction:

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Quantification of vector copy numbers and LM‐PCR Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

    Plasmid Preparation:

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Quantification of vector copy numbers and LM‐PCR Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

    Magnetic Cell Separation:

    Article Title: Heart non-specific effector CD4+ T cells protect from postinflammatory fibrosis and cardiac dysfunction in experimental autoimmune myocarditis
    Article Snippet: .. CD4+ T cell proliferation CD4+ T cells were isolated from mouse erythrocyte-lysed splenocytes/LNs or human PBMCs by MACS or sorted from hearts digested with Liberase (Roche). .. CD4+ T cell subpopulations were isolated using FACS Aria III.

    Software:

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome
    Article Snippet: .. Quantification of vector copy numbers and LM‐PCR Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software. .. VCN were determined on genomic DNA, isolated from the indicated tissues.

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    Roche human pbmc
    CAR T‐cell generation in human <t>PBMC</t> ‐transplanted mice Ex vivo generation of CAR T cells. Activated human PBMC were left untransduced or incubated with CD8‐LV CD19CAR at an MOI of 2. Five days later, expression of CD19‐CAR and CD8 was determined on CD3 + cells. Numbers indicate the percentage of cells in the respective gate. Experimental outline for in vivo CAR generation. 1 × 10 7 human PBMC were engrafted into naïve NSG mice or NSG mice that had been intraperitoneally (i.p.) injected with 5 × 10 5 Raji cells (Raji + ) 6 days before. One day later, 2 × 10 6 t.u. of CD8‐LV CD19CAR (filled circles) or CD8‐LV RFP (gray triangles) were i.p. injected, respectively. As further control, another group of mice received PBS (open circles). Seven days later, mice were sacrificed and organs and cells were removed for further analysis. Detection of CAR T cells by vector copy numbers <t>(VCN).</t> Genomic DNA was isolated from peritoneal cavity, spleen, and blood cells. VCN were determined in technical duplicates by qPCR for two individual mice of each group. The presence of B cells in the transplanted PBMC is indicated below. Cells isolated from the peritoneal cavity (peritoneum), spleen, or blood were evaluated by flow cytometry for the percentages of human CD8 + in CD3 + cells (D), of CAR + or RFP + cells in the CD8 + and CD8 − fractions, respectively (E), and of human CD19 + cells (F) within the fraction of human CD45 + cells. Representative density plots are shown for the peritoneal cells. The gating strategy is represented in Appendix Fig S1A . Mice were transplanted with B‐cell‐depleted human PBMC and then received CD8‐LV CD19CAR (filled circle) or PBS (open circle). As control, CD8‐LV CD19CAR or PBS was injected into mice transplanted with non‐depleted PBMC. Data information: Data represent mean ± SD for all groups (CD8‐LV CD19CAR : n = 6 (+Raji) and n = 4 (−Raji) in (D), n = 4 (−B‐cells) in (G); CD8‐LV RFP : n = 4; PBS: n = 4 in (G), all others n = 3). Statistical significance was determined using Mann–Whitney test; ns, not significant.
    Human Pbmc, supplied by Roche, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human pbmc/product/Roche
    Average 93 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    human pbmc - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    92
    Roche human peripheral blood mononuclear cells pbmc
    CAR T‐cell generation in human <t>PBMC</t> ‐transplanted mice Ex vivo generation of CAR T cells. Activated human PBMC were left untransduced or incubated with CD8‐LV CD19CAR at an MOI of 2. Five days later, expression of CD19‐CAR and CD8 was determined on CD3 + cells. Numbers indicate the percentage of cells in the respective gate. Experimental outline for in vivo CAR generation. 1 × 10 7 human PBMC were engrafted into naïve NSG mice or NSG mice that had been intraperitoneally (i.p.) injected with 5 × 10 5 Raji cells (Raji + ) 6 days before. One day later, 2 × 10 6 t.u. of CD8‐LV CD19CAR (filled circles) or CD8‐LV RFP (gray triangles) were i.p. injected, respectively. As further control, another group of mice received PBS (open circles). Seven days later, mice were sacrificed and organs and cells were removed for further analysis. Detection of CAR T cells by vector copy numbers <t>(VCN).</t> Genomic DNA was isolated from peritoneal cavity, spleen, and blood cells. VCN were determined in technical duplicates by qPCR for two individual mice of each group. The presence of B cells in the transplanted PBMC is indicated below. Cells isolated from the peritoneal cavity (peritoneum), spleen, or blood were evaluated by flow cytometry for the percentages of human CD8 + in CD3 + cells (D), of CAR + or RFP + cells in the CD8 + and CD8 − fractions, respectively (E), and of human CD19 + cells (F) within the fraction of human CD45 + cells. Representative density plots are shown for the peritoneal cells. The gating strategy is represented in Appendix Fig S1A . Mice were transplanted with B‐cell‐depleted human PBMC and then received CD8‐LV CD19CAR (filled circle) or PBS (open circle). As control, CD8‐LV CD19CAR or PBS was injected into mice transplanted with non‐depleted PBMC. Data information: Data represent mean ± SD for all groups (CD8‐LV CD19CAR : n = 6 (+Raji) and n = 4 (−Raji) in (D), n = 4 (−B‐cells) in (G); CD8‐LV RFP : n = 4; PBS: n = 4 in (G), all others n = 3). Statistical significance was determined using Mann–Whitney test; ns, not significant.
    Human Peripheral Blood Mononuclear Cells Pbmc, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 1 article reviews
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    CAR T‐cell generation in human PBMC ‐transplanted mice Ex vivo generation of CAR T cells. Activated human PBMC were left untransduced or incubated with CD8‐LV CD19CAR at an MOI of 2. Five days later, expression of CD19‐CAR and CD8 was determined on CD3 + cells. Numbers indicate the percentage of cells in the respective gate. Experimental outline for in vivo CAR generation. 1 × 10 7 human PBMC were engrafted into naïve NSG mice or NSG mice that had been intraperitoneally (i.p.) injected with 5 × 10 5 Raji cells (Raji + ) 6 days before. One day later, 2 × 10 6 t.u. of CD8‐LV CD19CAR (filled circles) or CD8‐LV RFP (gray triangles) were i.p. injected, respectively. As further control, another group of mice received PBS (open circles). Seven days later, mice were sacrificed and organs and cells were removed for further analysis. Detection of CAR T cells by vector copy numbers (VCN). Genomic DNA was isolated from peritoneal cavity, spleen, and blood cells. VCN were determined in technical duplicates by qPCR for two individual mice of each group. The presence of B cells in the transplanted PBMC is indicated below. Cells isolated from the peritoneal cavity (peritoneum), spleen, or blood were evaluated by flow cytometry for the percentages of human CD8 + in CD3 + cells (D), of CAR + or RFP + cells in the CD8 + and CD8 − fractions, respectively (E), and of human CD19 + cells (F) within the fraction of human CD45 + cells. Representative density plots are shown for the peritoneal cells. The gating strategy is represented in Appendix Fig S1A . Mice were transplanted with B‐cell‐depleted human PBMC and then received CD8‐LV CD19CAR (filled circle) or PBS (open circle). As control, CD8‐LV CD19CAR or PBS was injected into mice transplanted with non‐depleted PBMC. Data information: Data represent mean ± SD for all groups (CD8‐LV CD19CAR : n = 6 (+Raji) and n = 4 (−Raji) in (D), n = 4 (−B‐cells) in (G); CD8‐LV RFP : n = 4; PBS: n = 4 in (G), all others n = 3). Statistical significance was determined using Mann–Whitney test; ns, not significant.

    Journal: EMBO Molecular Medicine

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome

    doi: 10.15252/emmm.201809158

    Figure Lengend Snippet: CAR T‐cell generation in human PBMC ‐transplanted mice Ex vivo generation of CAR T cells. Activated human PBMC were left untransduced or incubated with CD8‐LV CD19CAR at an MOI of 2. Five days later, expression of CD19‐CAR and CD8 was determined on CD3 + cells. Numbers indicate the percentage of cells in the respective gate. Experimental outline for in vivo CAR generation. 1 × 10 7 human PBMC were engrafted into naïve NSG mice or NSG mice that had been intraperitoneally (i.p.) injected with 5 × 10 5 Raji cells (Raji + ) 6 days before. One day later, 2 × 10 6 t.u. of CD8‐LV CD19CAR (filled circles) or CD8‐LV RFP (gray triangles) were i.p. injected, respectively. As further control, another group of mice received PBS (open circles). Seven days later, mice were sacrificed and organs and cells were removed for further analysis. Detection of CAR T cells by vector copy numbers (VCN). Genomic DNA was isolated from peritoneal cavity, spleen, and blood cells. VCN were determined in technical duplicates by qPCR for two individual mice of each group. The presence of B cells in the transplanted PBMC is indicated below. Cells isolated from the peritoneal cavity (peritoneum), spleen, or blood were evaluated by flow cytometry for the percentages of human CD8 + in CD3 + cells (D), of CAR + or RFP + cells in the CD8 + and CD8 − fractions, respectively (E), and of human CD19 + cells (F) within the fraction of human CD45 + cells. Representative density plots are shown for the peritoneal cells. The gating strategy is represented in Appendix Fig S1A . Mice were transplanted with B‐cell‐depleted human PBMC and then received CD8‐LV CD19CAR (filled circle) or PBS (open circle). As control, CD8‐LV CD19CAR or PBS was injected into mice transplanted with non‐depleted PBMC. Data information: Data represent mean ± SD for all groups (CD8‐LV CD19CAR : n = 6 (+Raji) and n = 4 (−Raji) in (D), n = 4 (−B‐cells) in (G); CD8‐LV RFP : n = 4; PBS: n = 4 in (G), all others n = 3). Statistical significance was determined using Mann–Whitney test; ns, not significant.

    Article Snippet: Quantification of vector copy numbers and LM‐PCR Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software.

    Techniques: Mouse Assay, Ex Vivo, Incubation, Expressing, In Vivo, Injection, Plasmid Preparation, Isolation, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry, MANN-WHITNEY

    Clonality and exhaustion of in vivo ‐generated CAR T cells Clonality analysis by amplifying vector sequences integrated in genomic DNA. LM‐PCR detecting the integrated vector in genomic DNA purified from peritoneal and spleen cells harvested from PBMC‐transplanted mice injected with CD8‐LV CD19CAR (CAR) in the presence or absence of B cells or CD8‐LV RFP (RFP). Two mice of each group were analyzed. The internal control band is indicated as well as primer dimers in the water control (H 2 O). Cells isolated from peritoneal cavity or spleen of human PBMC‐transplanted NSG mice (± i.p. transplanted Raji cells) treated with CD8‐LV CD19CAR (CAR) or CD8‐LV RFP (RFP) were analyzed for expression of exhaustion markers by flow cytometry. CD8 + cells from viable human CD3 + cells were gated for transgene‐positive (CAR + or RFP + ) and transgene‐negative (CAR − or RFP − ) cells. These two cell populations were then separately gated for expression of PD‐1, LAG‐3, and TIM‐3. For the four experimental groups, percentages of positive cells for each exhaustion marker are shown for transgene‐positive and transgene‐negative CD8 + cells. Mean values ± SD are shown. N = 3 in samples with closed circles, n = 4 in samples with open circles or closed triangles, while for samples with open triangles, n = 3 for peritoneum and n = 4 for spleen. Statistical evaluation of the data was performed using one‐way ANOVA test with Bonferroni correction.

    Journal: EMBO Molecular Medicine

    Article Title: In vivo generation of human CD19‐ CAR T cells results in B‐cell depletion and signs of cytokine release syndrome

    doi: 10.15252/emmm.201809158

    Figure Lengend Snippet: Clonality and exhaustion of in vivo ‐generated CAR T cells Clonality analysis by amplifying vector sequences integrated in genomic DNA. LM‐PCR detecting the integrated vector in genomic DNA purified from peritoneal and spleen cells harvested from PBMC‐transplanted mice injected with CD8‐LV CD19CAR (CAR) in the presence or absence of B cells or CD8‐LV RFP (RFP). Two mice of each group were analyzed. The internal control band is indicated as well as primer dimers in the water control (H 2 O). Cells isolated from peritoneal cavity or spleen of human PBMC‐transplanted NSG mice (± i.p. transplanted Raji cells) treated with CD8‐LV CD19CAR (CAR) or CD8‐LV RFP (RFP) were analyzed for expression of exhaustion markers by flow cytometry. CD8 + cells from viable human CD3 + cells were gated for transgene‐positive (CAR + or RFP + ) and transgene‐negative (CAR − or RFP − ) cells. These two cell populations were then separately gated for expression of PD‐1, LAG‐3, and TIM‐3. For the four experimental groups, percentages of positive cells for each exhaustion marker are shown for transgene‐positive and transgene‐negative CD8 + cells. Mean values ± SD are shown. N = 3 in samples with closed circles, n = 4 in samples with open circles or closed triangles, while for samples with open triangles, n = 3 for peritoneum and n = 4 for spleen. Statistical evaluation of the data was performed using one‐way ANOVA test with Bonferroni correction.

    Article Snippet: Quantification of vector copy numbers and LM‐PCR Vector copy number (VCN) analysis of mice engrafted with human PBMC was performed by TaqMan‐based qPCR using a LightCycler® 480 Instrument II (Roche), and data were analyzed with LightCycler® Software.

    Techniques: In Vivo, Generated, Plasmid Preparation, Polymerase Chain Reaction, Purification, Mouse Assay, Injection, Isolation, Expressing, Flow Cytometry, Cytometry, Marker

    Viral loads, HBV antigens, and antibodies in coculture of HepG2.2.15 with IL-33-stimulated PBMCs. HepG2.2.15 cells were cocultured alone, treated with IL-33 (1 ng/mL), cocultured with PBMCs, IL-33, and PBMCs or IL-33/PBMCs/anti-IL-21, respectively.

    Journal: Journal of Interferon & Cytokine Research

    Article Title: IL-33 Enhances Humoral Immunity Against Chronic HBV Infection Through Activating CD4+CXCR5+ TFH Cells

    doi: 10.1089/jir.2013.0122

    Figure Lengend Snippet: Viral loads, HBV antigens, and antibodies in coculture of HepG2.2.15 with IL-33-stimulated PBMCs. HepG2.2.15 cells were cocultured alone, treated with IL-33 (1 ng/mL), cocultured with PBMCs, IL-33, and PBMCs or IL-33/PBMCs/anti-IL-21, respectively.

    Article Snippet: The concentrations of HBV DNA were examined in the serum samples from HBV-Tg mice, and the supernatants of HepG2.2.15 cells cocultured with human PBMCs were quantified by luciferase assays using the luciferase quantization detection kit (Roche Amplicor, Basel, Switzerland), according to the manufacturers' instruction (Jiang and others ).

    Techniques: