Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Schematic representation of M. pneumoniae M129 P1 gene and its four gene fragments; P1-I, P1-II, P1-III and P1-IV. Each bar represents the position of UGA codons that codes for tryptophan. To express these fragments, UGA codons were modified to UGG. Fragments were amplified using a set of forward (F) and reverse primers (R).

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Modification, Amplification

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: SDS-PAGE and Western blot analysis of recombinant M. pneumoniae P1 proteins fragments. (A) Coomassie blue stained SDS-PAGE analysis of rP1-I, rP1-II, rP1-III and rP1-IV in E. coli extract. The fragments were expressed in pET28b vector and protein production was induced with IPTG in E. coli. (B) Western blot analysis of induced and uninduced P1 protein fragments rP1-I, rP1-II, rP1-IV (i) and rP1-III (ii), showing reactivity with anti-6X His antibody. (C) Coomassie blue stained SDS-PAGE analysis of Ni 2+ -NTA purified P1 protein fragments; rP1-I, rP1-II, rP1-III and rP1-IV. (D) Western blot analysis of purified P1 protein fragments rP1-I, rP1-II, rP1-III and rP1-IV showing reactivity with anti-6X His antibody. Lane Marker: Molecular mass marker (kDa); Arrows indicate position of expressed protein.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: SDS Page, Western Blot, Recombinant, Staining, Plasmid Preparation, Purification, Marker

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Western blot and ELISA analysis of M. pneumoniae lysate and Cross reactivity of Pab (rP1-I) and Pab (rP1-IV). Reactivity of P1 (170 kDa) with anti-P1 protein fragments antibody Pab (rP1-I), Pab (rP1-II), Pab (rP1-III) & Pab (rP1-IV) rose in Rabbit by western blotting (A) and by ELISA (B) . (C) & (D) Immuno blot analysis of rP1-I, rP1-II, rP1-III and rP1-IV fragments with Pab (rP1-I) and Pab (rP1-IV) showing their cross reactivity with respective sera. Lane Marker: Molecular mass marker (kDa).

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Marker

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Recombinant P1 protein fragments are recognized by anti- M. pneumoniae antibody and by sera of M. pneumoniae infected patients. (A) (I) Coomassie blue stained SDS-PAGE analysis of purified M. pneumoniae P1 protein fragments; rP1-I, rP1-II, rP1-III and rP1-IV. Immuno blot analysis of purified P1 protein fragments; rP1-I, rP1-II, rP1-III and rP1-IV using anti- M. pneumoniae antibody (II) and using pooled sera of M. pneumoniae infected patients (III). (B) Immuno blot analysis of purified M. pneumoniae P1 protein fragments rP1-I, rP1-II, rP1-III and rP1-IV with several sera of M. pneumoniae infected patients. PM: Prestained protein marker; PC: positive control; NC: Negative control; Numbers over the blot indicate serial number of sera of M. pneumoniae infected patients tested for these experiments.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Recombinant, Infection, Staining, SDS Page, Purification, Marker, Positive Control, Negative Control

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Comparative ELISA analysis of recombinant P1 protein fragments with sera of M. pneumoniae infected patients. Reactivity of purified M. pneumoniae P1 proteins fragments with 25 sera of M. pneumoniae infected patients by ELISA (A) , with 16 healthy patient sera (B) and average values of both A & B (C) . Number on top of column indicates serial number of sera of M. pneumoniae infected patients tested for these experiments.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Infection, Purification

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: IFM adhesion assay of M. pneumoniae (A-E). The M. pneumoniae attached to the HEp-2 cells were detected by either anti- M. pneumoniae antibody or antibodies rose in rabbits. The detecting antibodies were added after fixation with methanol. (A) anti- M. pneumoniae antibody (positive control), (B) Pab (rP1-I), (C) Pab (rP1-II), (D) Pab (rP1-III), (E) Pab (rP1-IV). IFM surface exposure assay of M. pneumoniae (F-J) . In this assay the detecting antibodies were added before the methanol fixation. (F) anti- M. pneumoniae antibody (positive control), (G) Pab (rP1-I), (H) Pab (rP1-II), (I) Pab (rP1-III), (J) Pab (rP1-IV). Negative controls: (K) mycoplasmas alone (Without Pabs), (L) Pabs alone (Without mycoplasmas). Bar, 2 μm.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Cell Adhesion Assay, Positive Control

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: IFM adhesion inhibition assay. M. pneumoniae were pre-incubated with either anti- M. pneumoniae antibodies or antibodies rose in rabbits in different dilutions (1:50, 1:100, 1:200, 1:500) before infection of the HEp-2 cells. These antibodies were: (A-D) anti- M. pneumoniae antibody (positive control), (E-H) Pab (rP1-I), (I-L) Pab (rP1-IV), (M) Pab (rP1-II) (N) Pab (rP1-III) (O) Without antibody, (P) pre-immune serum. Bar, 2 μm.

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Inhibition, Incubation, Infection, Positive Control

Journal: BMC Microbiology

Article Title: Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

doi: 10.1186/1471-2180-14-108

Figure Lengend Snippet: Primer sequence used to amplify all four fragments of M. pneumoniae M129 P1 gene

Article Snippet: Entire M. pneumoniae M129 P1 gene was synthesized in four fragments; N-terminal P1-I (1069 bp), two middle fragments P1-II (1043 bp) and P1-III (1983 bp), and C-terminal P1-IV (1167 bp) fragments by codon optimization replacing 21 UGA to UGG codons (Entelechon GmbH, Germany).

Techniques: Sequencing

Journal: Transfusion

Article Title: Convalescent plasma treatment of critically ill intensive care COVID ‐19 patients

doi: 10.1111/trf.16392

Figure Lengend Snippet: SARS‐CoV‐2 IgG ELISA (Euroimmun) ratios in sera of convalescent volunteers and in control samples. In total, 61 sera from convalescent, potential blood donors with a SARS‐CoV‐2 PCR‐confirmed infection (left panel) and in twenty 3‐year‐old retention samples from our blood bank (right panel) were tested for SARS‐CoV‐2 spike S1 domain–specific IgG. Samples were classified as positive (OD Ratio ≥ 1.1), borderline (OD Ratio ≥ 0.8 to <1.1), or negative (OD Ratio < 0.8). Horizontal bars represent the median and the interquartile range. The gray shaded area indicates borderline results

Article Snippet: SARS‐CoV‐2–specific antibodies were determined by an enzyme‐linked immunosorbent assay detecting immunoglobulin G (IgG) antibodies against the S1 protein (Euroimmun), and the neutralizing titers were determined with a cell‐culture‐based neutralization assay.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Infection

Journal: Transfusion

Article Title: Convalescent plasma treatment of critically ill intensive care COVID ‐19 patients

doi: 10.1111/trf.16392

Figure Lengend Snippet: Distribution of antibody ratios in sera of convalescent volunteers acquired at different periods after the disease onset. SARS‐CoV‐2 spike S1 antibodies were measured by ELISA (Euroimmun) in sera from convalescent, potential blood donors with PCR‐confirmed SARS‐CoV‐2 infection. Correlation analysis was performed by Spearman's test. The continuous line shows the regression line, and the broken lines show the 95% confidence interval

Article Snippet: SARS‐CoV‐2–specific antibodies were determined by an enzyme‐linked immunosorbent assay detecting immunoglobulin G (IgG) antibodies against the S1 protein (Euroimmun), and the neutralizing titers were determined with a cell‐culture‐based neutralization assay.

Techniques: Enzyme-linked Immunosorbent Assay, Infection