Product Code IF1250M Rev. I Performance Characteristics Not for Distribution in the United States EXPECTED VALUES Community Acquired Pneumonia Population Two outside investigators assessed the Focus Chlamydia MIF IgM and the Focus Chlamydia MIF IgG ("Focus MIFs") with a total of 144 community acquired pneumonia patients. 141 of the 144 were sequentially selected, and three patients were selected because they were PCR positive for C. pneumoniae. Each patient was an adult outpatient, and their pneumonia was confirmed with a positive chest x-ray. Serum samples were drawn twice, the first serum was drawn 2 weeks or more postonset of symptoms, and the second serum was drawn 3 to 6 weeks after the first. Each patient was tested for C. pneumoniae, Legionella pneumophila, Mycoplasma pneumoniae and Streptococcus pneumoniae. C. pneumoniae testing included culture, PCR, and reference MIFs (IgG and IgM) from a university in the Pacific Northwest. Legionella pneumophila testing included serology (polyvalent antibody), urinary antigen and PCR. Mycoplasma pneumoniae testing included culture, PCR and serology (IgG and IgM). Streptococcus pneumoniae testing included urinary antigen. The observed prevalences and the hypothetical predictive values for the population are shown in the tables below. Community Acquired Pneumonia Outpatients Prevalence vs. Pathogen Prevalence C. pneumoniae (presumed and possible acute) 12.8% C. pneumoniae (presumed acute) 3.5% C. pneumoniae (possible acute) 9.2% Undifferentiated Chlamydia (presumed acute) 0.7% Legionella pneumophila 2.8% Mycoplasma pneumoniae 13.5% Streptococcus pneumoniae 8.5% Unidentified Pathogen, including C. trachomatis (past infection) C. trachomatis (possible acute) C. psittaci (past infection) 61.0% PERFORMANCE CHARACTERISTICS Reactivity with Community Acquired Pneumonia Patients Two outside investigators evaluated the Focus MIF IgM and IgG (Focus MIFs) reactivity with a total of 144 community acquired pneumonia (CAP) patients. The patients and test methods are described in the EXPECTED VALUES section (above). Focus MIF reactivity was calculated for the IgM MIF alone, the IgG MIF alone and the combined reactivity of both of the Focus MIF IgM and Focus MIF IgG ("Combined Focus MIFs"). Predictive values are based on the estimated reactivity of both of the Focus MIF IgM and Focus MIF IgG combined ("Combined Focus MIFs") for C. pneumoniae: 99.8% (85/86) specificity with the combined Focus MIFs, 87.5% (7/8) sensitivity with the combined Focus MIFs, 37.5% (3/8) sensitivity with the Focus MIF IgM, and 62.5% (5/8) sensitivity with the Focus MIF IgG. 99.8% (85/86) specificity with the Focus MIF IgG. Prevalence vs. Hypothetical Predictive Values for C. pneumoniae Pos Prev Combined IgM IgG % PPV % NPV % PPV % NPV % PPV % NPV % 30 99.5 94.9 100 78.9 99.3 86.1 25 99.3 96.0 100 82.8 99.0 88.9 20 99.1 97.0 100 86.5 98.7 91.4 15 98.7 97.8 100 90.1 98.2 93.8 10 98.0 98.6 100 93.5 97.2 96.0 5 95.8 99.3 100 96.8 94.3 98.1 1 81.5 99.9 100 99.4 75.9 99.6
Page 2 The unidentified pathogen pneumonia group (negative for all tests performed) included 86 of 144 patients. Focus MIF reactivity for C. pneumoniae with patients infected with unidentified pathogens was as follows: 99.8% (85/86) specificity with the combined Focus MIFs, 99.8% (85/86) specificity with the Focus MIF IgG. Focus MIF reactivity for C. trachomatis with patients infected with unidentified pathogens was as follows: 100% (86/86) specificity with the combined Focus MIFs, 100% (86/86) specificity with the Focus MIF IgG (there were two past C. trachomatis infections detected). The C. pneumoniae presumed acute pneumonia group (PCR positive, and/or IgM reference MIF positive, and/or reference MIF IgG with a 4-fold rise) included 8 of 144 patients. Focus MIF reactivity for C. pneumoniae with presumed acute C. pneumoniae patients was as follows: 87.5% (7/8)* sensitivity with the combined Focus MIFs, 37.5% (3/8) sensitivity with the Focus MIF IgM, and 62.5% (5/8)* sensitivity with the Focus MIF IgG. * The one Focus C. pneumoniae negative sample was detected by the Focus IgG MIF as an undifferentiated Chlamydia presumed acute infection (C. trachomatis positive and C. psittaci positve). Focus MIF reactivity for C. trachomatis with presumed acute C. pneumoniae patients was as follows: 100% (7/7)** specificity with the combined Focus MIFs, 100% (8/8) specificity with the Focus MIF IgM, and 100% (7/7)** specificity with the Focus MIF IgG. ** Excludes one Focus C. pneumoniae negative sample that was detected by the Focus IgG MIF as an undifferentiated Chlamydia presumed acute infection (C. trachomatis positive and C. psittaci positve). The C. pneumoniae possible acute pneumonia group (reference MIF IgM negative, culture negative, PCR negative, and with a reference MIF IgG endpoint of 1:512 or greater) included 13 of 144 patients. Focus MIF reactivity for C. pneumoniae with possible acute C. pneumoniae patients was as follows: 100% (13/13) agreement between the Focus MIF IgM and reference IgM MIF (both IgM MIFs were negative), and 23.1% (3/13) sensitivity with the Focus MIF IgG. The presumed Legionella pneumophila pneumonia group (Legionella sero-positive) included 3 of 144 patients. Focus MIF reactivity for C. pneumoniae with presumed Legionella pneumophila patients was as follows: 100% (3/3) specificity with the combined Focus MIFs, 100% (3/3) specificity with the Focus MIF IgM, and 100% (3/3) specificity with the Focus MIF IgG. Focus MIF reactivity for C. trachomatis with presumed Legionella pneumophila patients was as follows: 100% (3/3) specificity with the combined Focus MIFs, 100% (3/3) specificity with the Focus MIF IgM, and 100% (3/3) specificity with the Focus MIF IgG. The presumed Mycoplasma pneumoniae pneumonia group (Mycoplasma IgM positive and/or Mycoplasma IgG with a 4-fold rise) included 19 of 144 patients. One of the 19 patients was positive for both Mycoplasma pneumoniae and Streptococcus pneumoniae. Focus MIF reactivity for C. pneumoniae with presumed Mycoplasma pneumoniae patients was as follows: 100% (19/19) specificity with the combined Focus MIFs, 100% (19/19) specificity with the Focus MIF IgM, and 100% (19/19) specificity with the Focus MIF IgG. Focus MIF reactivity for C. trachomatis with presumed Mycoplasma pneumoniae patients was as follows: 100% (19/19) specificity with the combined Focus MIFs, 100% (19/19) specificity with the Focus MIF IgM, and 100% (19/19) specificity with the Focus MIF IgG. The presumed Streptococcus pneumoniae pneumonia group (Streptococcus pneumoniae urinary antigen positive) included 12 of 144 patients. One of the 12 patients was positive for both Mycoplasma pneumoniae and Streptococcus pneumoniae. Focus MIF reactivity for C. pneumoniae with presumed Streptococcus pneumoniae patients was as follows: 100% (12/12) specificity with the combined Focus MIFs, 100% (12/12) specificity with the Focus MIF IgM, and 100% (12/12) specificity with the Focus MIF IgG. Focus MIF reactivity for C. trachomatis with presumed Streptococcus pneumoniae patients was as follows: 100% (12/12) specificity with the combined Focus MIFs, 100% (12/12) specificity with the Focus MIF IgM, and 100% (12/12) specificity with the Focus MIF IgG.
Page 3 Focus MIF % C. pneumoniae Positive 100 90 80 70 60 50 40 30 20 10 0 C. pneumo L. pneumo M. pneumo Pathogen S. pneumo IgM IgG Combined Unidentified Focus MIF Reactivity to C. pneumoniae with Community Acquired Pneumonia Patients Community Acquired Pneumonia Patients (C. pneumoniae) Focus MIF % C. pneumoniae Positive IgM IgG Combined (IgG+IgM) Unidentified Pathogen 0.0% (0/86) 1.2% (1/86) 1.2% (1/86) 95%CI 0.0-6.3% 95%CI 0.0-6.3% C. pneumoniae (Presumed Acute) 37.5% (3/8) 62.5% (5/8) 87.5% (7/8) 95%CI 8.5-75.5% 95%CI 24.5-91.5% 95%CI 47.3-99.7% C. pneumoniae (Possible Acute) 0.0% (0/13) 23.1% (3/13) 23.1% (3/13) 95%CI 5.0-53.8% 95%CI 5.0-53.8% C. pneumoniae (Possible Acute) & L. pneumophila 75.0% (3/4)* 95%CI 28.4-99.5% 75.0% (3/4)* 95%CI 28.4-99.5% Legionella pneumophila Mycoplasma pneumoniae Streptococcus pneumoniae * 2/3 of the Focus IgG positives were possible acutes ( 512), and the third positive was a presumed acute ( 4X rise). Includes one sample that was positive for Streptococcus pneumoniae and Mycoplasma pneumoniae The one Focus IgG that was C pneumoniae positive in the unidentified pathogen group was a presumed acute ( 4X rise). Community Acquired Pneumonia Patients Focus MIF C. trachomatis % Positive IgM IgG Combined (IgG+IgM) C. pneumoniae (Presumed Acute) C. pneumoniae (Possible Acute) C. pneumoniae (Possible Acute) & L. pneumophila Legionella pneumophila Mycoplasma pneumoniae Streptococcus pneumoniae Unidentified Pathogen 0.0% (0/8) 95%CI 0.0-36.9% 0.0% (0/13) 0.0% (0/86) 0.0% (0/7)* 95%CI 0.0-41.0% 0.0% (0/13) 0.0% (0/86) 0.0% (0/7) 95%CI 0.0-41.0% 0.0% (0/13) 0.0% (0/86) * Excludes one presumed acute C. pneumoniae sample that was an undifferentiated Chlamydia positive (C. trachomatis and C. psittaci positive) in the Focus MIF which may have not been identified in the reference MIF. Two past C. trachomatis infections were detected in each of the possible C pneumoniae infection group and the unidentified pathogen group.
Page 4 Chlamydia pneumoniae Culture/PCR Positives An outside investigator evaluated the test's reactivity with 98 sera from 27 patients that exhibited symptoms of acute respiratory disease. Each patient was Chlamydia pneumoniae culture and/or PCR positive population. The Focus IgM MIF was C. pneumoniae positive for 68.4% (67/98) of sera, and C. pneumoniae positive for 88.9% (24/27) of patients. The Focus IgM MIF was C. trachomatis positive for 0.0% (0/98) of sera, and C. trachomatis positive for 0.0% (0/27) of patients. C. pneumoniae Culture/PCR Positives C. pneumoniae culture/pcr positives (reactivity by sera) 68.4% (67/98) 95% CI 58.2-77.4% 0.0% (0/98) 95% CI 0.0-3.7% C. pneumoniae culture/pcr positives (reactivity by patient) 88.9% (24/27) 95% CI 70.8-97.7% 0.0% (0/27) 95% CI.0-12.8% * Sera from early draws (2-3 weeks from onset) from 2 patients were undifferentiated Chlamydia positive (positive on all 3 antigen spots) by the Focus assay. C. trachomatis Culture Positives An outside investigator evaluated the test's cross-reactivity with 75 sera from 29 patients that were C. trachomatis culture positive. Most patients (27/29) were infants. 35 sera were C. trachomatis seropositive, including 2 having endpoints of 1:1280. The Focus IgM MIF was C. pneumoniae positive with 1.3% (1/75) of sera, 3.5% (1/29) of patients, and 0.0% (0/35) of reference MIF C. trachomatis sero-positives. The Focus IgM MIF was C. trachomatis positive with 46.7% (35/75) of sera, and 75.9% (22/29) of patients. Reactivity with C. trachomatis Culture Positives C. trachomatis culture positives (reactivity by sera) 1.3% (1/75) 95% CI 0.0-7.2% 46.7% (35/75) 95% CI 35.0-58.6% C. trachomatis culture positives (reactivity by patient) 3.5% (1/29) 95% CI 0.0-17.8% 75.9% (22/29) 95% CI 56.5-89.7% Reactivity with C. psittaci Seropositives An outside investigator evaluated the Focus MIF IgM's cross-reactivity with 42 sera from 19 Chlamydia psittaci sero-positive patients. All of the patients were seropositive by Chlamydia complement fixation, and sero-positive by a MIF (IgM and/or IgG) from a university located in the Pacific Northwest United States, and each patient had epidemiological evidence of Chlamydia psittaci infection. 16 of the 19 patients were drawn more than once. The Focus IgM MIF was C. pneumoniae positive with 10.5% (2/19) of the patients, and 11.9% (5/42) of the sera. The Focus IgM MIF was C. trachomatis positive with was 10.5% (2/19) of the patients, and 7.1% (3/42) of the sera. The Focus IgM MIF was C. psittaci positive with 57.9% (11/19) of the patients, and 52.4% (22/42) of the sera. Reactivity with C. psittaci Seropositives 11.9% (5/42)* 7.1% (3/42)** C. psittaci seropositives (reactivity by sera) 95% CI 4.0-25.6% 95% CI 1.5-19.5% C. psittaci seropositives (reactivity by 10.5% (2/19) 10.5% (2/19) patient) 95% CI 1.3-33.1% 95% CI 1.3-33.1% * 4/5 of the positives were undifferentiated Chlamydia positives (positive with both of the C. psittaci and C. pneumoniae antigen spots). ** 3/3 of the positives were undifferentiated Chlamydia positives (positive with two or more of the antigen spots: the C. trachomatis and either the C. psittaci and/or C. pneumoniae antigen spots). Reactivity with Blood Donors An outside investigator evaluated the test's reactivity with sera from 100 blood donors. The Focus IgM MIF was C. pneumoniae positive with 2.0% (2/100) and C. trachomatis positive for 1.0% (1/100) of blood donors. The C. pneumoniae and C. trachomatis results both include one serum that was an undifferentiated Chlamydia positive (positive with the C trachomatis and C. pneumoniae antigen spots by the Focus MIF). Reactivity with Blood Donors (n = 100) Blood Donors 2.0% (2/100)* 95% CI 0.2-7.0% 1.0% (1/100)* 95% CI 0.0-5.5% * Includes 1 serum that was an undifferentiated Chlamydia positive (positive with the C trachomatis and C. pneumoniae antigen spots) by the Focus assay.
Page 5 Cross-reactivity with Microbes Causing Similar Symptoms An investigator evaluated the test's reactivity with Mycoplasma pneumoniae IgM sero-positives and Legionella pneumophila sero-positives The Focus IgM MIF was C. pneumoniae positive with 0.0% (0/10) of M. pneumoniae IgM sero-positives, and 0.0% (0/5) of L. pneumophila sero-positives. The Focus IgM MIF was also C. trachomatis negative with all of the M. pneumoniae samples and all of the L. pneumophila samples. Cross-reactivity with Microbes Causing Similar Symptoms Mycoplasma pneumoniae IgM sero-positives 0.0% (0/10) 95% CI 0.0-30.8% Legionella pneumophila sero-positives 0.0% (0/5) 0.0% (0/10) 95% CI 0.0-30.8% 0.0% (0/5) Cross-reactivity with Common Viruses An investigator evaluated the test's reactivity with EBV (Viral Capsid Antigen) IgM positives, CMV IgM positives and Influenzae A IgG positives. The Focus IgM MIF was C. pneumoniae positive with of EBV (Viral Capsid Antigen) IgM positives, 0.0% (0/11) of CMV IgM positives, and 0.0% (0/5) of Influenzae A IgG positives. The Focus IgM MIF was also C. trachomatis negative with all of the EBV, CMV and Influenzae A samples. Cross-reactivity with Common Viruses EBV VCA IgMo seropositives 95% CI 0.0-70.8% CMV IgM seropositives 0.0% (0/11) 95% CI 0.0-28.5% Influenzae A IgG seropositives 0.0% (0/5) 95% CI 0.0-70.8% 0.0% (0/11) 95% CI 0.0-28.5% 0.0% (0/5) Interference with Rheumatoid Factor & Anti-Nuclear Antibody An investigator evaluated interference with six rheumatoid factor sera and six anti-nuclear antibody sero-positives. The Focus IgM MIF was C. pneumoniae positive with 0.0% (0/6) of rheumatoid factor sera, and 0.0% (0/6) of anti-nuclear antibody sero-positives. The Focus IgM MIF was also C. trachomatis negative with all of the rheumatoid factor sera, and all of the anti-nuclear antibody sera. Interference with Rheumatoid Factor & Anti-Nuclear Antibody % Positives by sera 0.0% (0/6) 0.0% (0/6) % Positives by patient 0.0% (0/6) 0.0% (0/6) IgG removal by IgM Pretreatment Diluent An investigator evaluated IgG removal by pretreatment diluent using six samples with IgG 700mg/dL. IgG concentration was determined by nephelometry before and after pretreatment. The Pretreatment reduced IgG at least 100-fold on all six samples. Inter-assay Reproducibility An investigator evaluated inter-assay reproducibility by testing 24 sera endpointed with all three antigens, in 3 runs, with one run on three separate days. 99% (213/216) of endpoints were within 2-fold dilution among all repeated runs. Inter-lot Reproducibility An investigator evaluated inter-lot reproducibility by testing 15 sera endpointed with all three antigens with three different lots of slides. 100% (135/135) of endpoints were within 2-fold dilution among all repeated runs. Inter-laboratory Reproducibility Two off-site investigators evaluated inter-laboratory reproducibility by testing 15 sera endpointed with all three antigens. 98% (265/270) of endpoints were within 2- fold dilution among all 6 runs. AUTHORIZED REPRESENTATIVE mdi Europa GmbH, Langenhagener Str. 71, 30855 Langenhagen-Hannover, Germany ORDERING INFORMATION Telephone: (562) 240-6500 (International) Fax: (562) 240-6510 PC.IF1250M Rev. I Date written: 27 Jan 2011 TECHNICAL ASSISTANCE Telephone: (562) 240-6500 (International) Fax: (562) 240-6526 Visit our web site at: www.focusdx.com Cypress, California 90630, U.S.A