Chlamydia MIF IgG. Performance Characteristics. Product Code IF1250G Rev. J. Not for Distribution in the United States
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1 Product Code IF1250G Rev. J 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 post-onset 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% (1.4%) (1.4%) (1.4%) 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 reactivities 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. 100% (86/86) specificity with the Focus MIF IgM, and 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 %
2 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, 100% (86/86) specificity with the Focus MIF IgM, and 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 IgM, and 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 positive). 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 positive). 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.
3 Page 3 Focus MIF % C. pneumoniae Positive C. pneumo L. pneumo M. pneumo Pathogen S. pneumo Unidentified Focus MIF Reactivity to C. pneumoniae with Community Acquired Pneumonia Patients Community Acquired Pneumonia Patients (C. pneumoniae) Focus MIF % C. pneumoniae Positive Unidentified Pathogen 0.0% (0/86) 1.2% (1/86) 1.2% (1/86) 95%CI % 95%CI % 95%CI % C. pneumoniae (Presumed Acute) 37.5% (3/8) 62.5% (5/8) 87.5% (7/8) 95%CI % 95%CI % 95%CI % C. pneumoniae (Possible Acute) 0.0% (0/13) 23.1% (3/13) 23.1% (3/13) 95%CI % 95%CI % 95%CI % C. pneumoniae (Possible Acute) & L. pneumophila 75.0% (3/4)* 95%CI % 75.0% (3/4)* 95%CI % Legionella pneumophila 95%CI % 95%CI % 95%CI % Mycoplasma pneumoniae 0.0% (0/19) 0.0% (0/19) 0.0% (0/19) 95%CI % 95%CI % 95%CI % Streptococcus pneumoniae 0.0% (0/12) 0.0% (0/12) 0.0% (0/12) 95%CI % 95%CI % 95%CI % * 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 (C. trachomatis) Focus MIF C. trachomatis % Positive C. pneumoniae (Presumed Acute) 0.0% (0/8) 0.0% (0/7)* 0.0% (0/7) 95%CI % 95%CI % 95%CI % C. pneumoniae (Possible Acute) 0.0% (0/13) 0.0% (0/13) 0.0% (0/13) 95%CI % 95%CI % 95%CI % C. pneumoniae (Possible Acute) & L. pneumophila Legionella pneumophila 95%CI % 95%CI % 95%CI % Mycoplasma pneumoniae 0.0% (0/19) 0.0% (0/19) 0.0% (0/19) 95%CI % 95%CI % 95%CI % Streptococcus pneumoniae 0.0% (0/12) 0.0% (0/12) 0.0% (0/12) 95%CI % 95%CI % 95%CI % Unidentified Pathogen 0.0% (0/86) 0.0% (0/86) 0.0% (0/86) 95%CI % 95%CI % 95%CI % * 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. IgM IgG Combined
4 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. Focus MIF reactivity for C. pneumoniae with C. pneumoniae culture positive patients was as follows: 92.6% (25/27) sensitivity with the combined Focus MIFs, 88.9% (24/27) sensitivity with the Focus MIF IgM, and 51.9% (14/27) sensitivity with the Focus MIF IgG. Focus MIF reactivity for C. trachomatis with C. pneumoniae culture positive patients was as follows: 100% (27/27) specificity with the combined Focus MIFs, 100% (27/27) specificity with the Focus MIF IgM, and 100% (27/27) specificity with the Focus MIF IgG. Reactivity with C. pneumoniae Culture/PCR Positives Focus MIF % Positive (by patient) Focus C. pneumoniae reactivity to C. pneumoniae PCR/culture positives Focus C. trachomatis reactivity to C. pneumoniae PCR/culture positives 88.9% (24/27) 95%CI % 100% (27/27) 95%CI % 51.9% (14/27) 95%CI % 100% (27/27) 95%CI % 92.6% (25/27) 95%CI % 100% (27/27) 95%CI % 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. Focus MIF reactivity for C. pneumoniae with Chlamydia trachomatis infantile pneumonia patients was as follows: 100% (27/27) specificity with the combined Focus MIFs, 100% (27/27) specificity with the Focus MIF IgM, and 100% (27/27) specificity with the Focus MIF IgG. Focus MIF reactivity for C. trachomatis with Chlamydia trachomatis infantile pneumonia patients was as follows: 85.2% (23/27) sensitivity with the combined Focus MIFs, 815% (22/27) sensitivity with the Focus MIF IgM, and 63.0% (17/27) sensitivity with the Focus MIF IgG. Reactivity with C. trachomatis Culture Positives Focus MIF % Positive* Focus C. pneumoniae reactivity to C. trachomatis culture positives 0.0% (0/27) 95%CI % 0.0% (0/27) 95%CI % 0.0% (0/27) 95%CI % Focus C. trachomatis reactivity to C. trachomatis culture positives 815% (22/27) 95%CI % 63.0% (17/27) 95%CI % 85.2% (23/27) 95%CI % * Positive if IgM positive or at least a 4x rise in IgG endpoint, or an IgG endpoint of 1:512 or greater. Reactivity with C. psittaci Seropositives An outside investigator evaluated the Focus MIF IgG'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 IgG MIF was C. pneumoniae positive (4x rise in paired sera or a single endpoint of >1:512) with 37.5% (6/16) of the patients, and 4/6 of the Focus positives were classified by the Focus MIF as undifferentiated chlamydia presumed acute infections (reactive to multiple chlamydia anigen spots. The Focus IgG MIF was C. trachomatis positive (4x rise in paired sera or a single endpoint of >1:512) with 6.3% (1/16) of the patients. The one Focus IgG MIF C. trachomatis positive was an undifferentiated chlamydia presumed acute positive (reactive to at least two chlamydia antigen spots). The Focus IgG MIF was C. psittaci positive (single serum endpoint 1:16) with 68.4% (13/19) of the patients, and 71.4% (30/42) of the sera. Reactivity with C. psittaci Seropositives Focus MIF IgG % Positive C. pneumoniae C. trachomatis C. psittaci seropositives (by patient) 37.5% (6/16)* 95% CI % 6.3% (1/16) 95% CI % * 4/6 of the C. pneumoniae positives and the one C. trachomatis positive were undifferentiated chlamydia positives (positive with multiple chlamydia antigen spots). Reactivity with Blood Donors An outside investigator evaluated the test's reactivity with sera from 64 blood donors. Resuts are for single serum only, and indicate reactivity only, or if the IgG endpoint was 1:512 or greater, then possible acute infection. Presumed acute infections (4x rise in IgG endpoint) can not be calculated from single serum results. The Focus IgG MIF was C. pneumoniae reactive with 42.2% (27/64) of the blood donors. 3 of the 27 had a Focus IgG MIF C. pneumoniae MIF endpoint of 1:512 or greater. The Focus IgG MIF was C. trachomatis reactive for 11.0% (7/64) of blood donors. None of samples had a Focus IgG MIF C. trachomatis MIF endpoint of 1:512 or greater. The Focus IgG MIF was C.psittaci reactive for 0.0% (0/64) of blood donors.
5 Reactivity with Blood Donors (n = 64) Focus MIF IgG % Reactive* Blood Donors 42.2% (27/64) 95%CI % C. pneumoniae MIF C. trachomatis MIF 11.0% (7/64) 95%CI % Chlamydia MIF IgG Page 5 * Single serum results only. 4x rise in endpoint could not be calculated. 3 samples had a Focus MIF IgG C. pneumoniae endpoint of 1:512 or greater. None of the samples had a Focus C. trachomatis endpoint of 1:512 or greater. Reactivity with Sexually Active Patients C. trachomatis An outside investigator in the midwestern United States compared the Focus Chlamydia MIF IgG to Kit A (a commercially available ELISA). Both kits were evaluated to determine the reactivity to Chlamydia trachomatis on a total of 400 sexually active patients. Three hundred of the patient samples were actively collected from a sexually transmitted disease (STD) clinic in the Pacific Northwest United States. One hundred of the patient samples were from an Australian laboratory collecting samples from a STD clinic that tested PCR positive for Chlamydia trachomatis. In cases where Focus Chlamydia MIF IgG and Kit A did not agree, the samples were tested on Kit B (a commercially available MIF). Percent agreements were calculated for Focus Chlamydia MIF IgG, Kit A and Kit B. Additionally, each population was analyzed separately and combined. STD Clinic Population PCR+ Population Focus MIF IgG Kit A (C. trachomatis) (C. trachomatis) + +/ N = 300 Positive Agreement: 37.3%, 95%CI % Negative Agreement: 92.6%, 95%CI % Total Agreement: 81.0%, 95%CI % Focus MIF IgG Kit A (C. trachomatis) (C. trachomatis) + +/ N = 100 Positive Agreement: 55.2%, 95%CI % Negative Agreement: 97.4%, 95%CI % Total Agreement: 70.0%, 95%CI % Total Population Kit A (C. trachomatis) Focus MIF IgG (C. trachomatis) + +/ N = 400 Positive Agreement: 45.9%, 95%CI % Negative Agreement: 93.2%, 95%CI % Total Agreement: 78.0%, 95%CI % A total of 88 samples did not agree between Focus MIF IgG and Kit A. All 88 samples were assayed on Kit B to referee the result with the exception of 2 samples (N=86*). The total agreement to Kit B for Focus MIF IgG and Kit A was 58.1% (50/86) and 36.0% (31/86), respectively. A total of 37 Focus MIF IgG negative samples were also negative on Kit B, 78.7% (37/47). A total of 7 Kit A negative samples were determined to be negative on Kit B, 14.9% (7/47). Thirty-nine samples tested positive on Kit B. Thirteen samples agreed and tested positive on Focus MIF IgG and Kit B. Twenty-four samples agreed and tested positive on Kit A and Kit B. As a result of referee testing, the positive agreement for Focus MIF IgG and Kit B was 33.3% (13/39). Focus MIF IgG (C. trachomatis) Kit B (C. trachomatis) Kit A (C. trachomatis) Kit B (C. trachomatis) / 3 7 N = 86* 12 7 Positive Agreement: 33.3%, 95%CI % N = 86* Negative Agreement: 78.7%, 95%CI % Positive Agreement: 61.5%, 95%CI % Total Agreement: 58.1%, 95%CI % Negative Agreement: 14.9%, 95%CI % Total Agreement: 36.0%, 95%CI % 2 samples were not included because tests on Kit B were not performed. Confirmed PCR+ Each product was also evaluated to determine the positive agreement to 100 (33 of the 100 samples were assayed by Kit B) patients that were from an Australian laboratory collecting samples from a STD clinic that tested PCR positive for Chlamydia trachomatis. Focus MIF IgG PCR N = 100 Positive Agreement: 32.0%, 95%CI % + Kit A PCR / 3 39 N = 100 Positive Agreement: 58.0%, 95%CI % + Kit B PCR N = 33 Positive Agreement: 51.5%, 95%CI % +
6 Page 6 CROSS REACTIVITY For those samples, which are negative by Kit B (MIF) for C. trachomatis and positive for either C. pneumoniae and/or C. psittaci, the cross-reactivity percent agreement with Focus MIF IgG and Kit A (ELISA) are calculated below. 93.3% (84/90): Focus MIF IgG cross-reactivity percent agreement to C. pneumoniae. 100% (90/90): Focus MIF IgG cross-reactivity percent agreement to C. psittaci. 80.0% (72/90): Kit A cross-reactivity percent agreement to C. pneumoniae. 100% (90/90): Kit A cross-reactivity percent agreement to C. psittaci. 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 IgG MIF was C. pneumoniae reactive with 0.0% (0/5) of EBV Viral Capsid Antigen IgG positives, 18.2% (2/11) of CMV IgM positives, and 0.0% (0/5) of influenzae A IgG positives. The Focus IgG MIF was also C. trachomatis negative with all of the EBV, CMV and influenzae A samples. Cross-reactivity with Common Viruses Focus MIF IgG % Positive C. pneumoniae MIF C. trachomatis MIF EBV VCA IgG seropositives 0.0% (0/5) 95% CI % CMV IgM seropositives 18.2% (2/11) 95% CI % Influenzae A IgG seropositives 0.0% (0/5) 95% CI % 0.0% (0/5) 95% CI % 0.0% (0/11) 95% CI % 0.0% (0/5) 95% CI % 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. 86% (186/216) of endpoints were within one 2-fold dilution. 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% (108/108) of endpoints were within one 2-fold dilution. Inter-laboratory Reproducibility Two off-site investigators evaluated inter-laboratory reproducibility by testing 15 sera endpointed with all three antigens. 89% (240/270) of endpoints were within one 2-fold dilution on all 6 runs. AUTHORIZED REPRESENTATIVE mdi Europa GmbH, Langenhagener Str. 71, Langenhagen-Hannover, Germany ORDERING INFORMATION Telephone: (562) (International) Fax: (562) TECHNICAL ASSISTANCE Telephone: (562) (International) Fax: (562) Visit our web site at: PC.IF1250G Rev. J Date written: 27 Jan 2011 Cypress, California 90630, U.S.A
Chlamydia MIF IgM. Performance Characteristics. Product Code IF1250M Rev. I. Not for Distribution in the United States
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