chlamydia culture sensitivity

Sensitivity and specificity estimates with 95% confidence intervals (CI) were reported as our main outcome measures. Terms and Conditions, 0000033708 00000 n the prevalence of chlamydial infection by culture was 4.60Zo (57/1241). When the first non-culture tests (antigen detection tests such as enzyme immunoassay (EIA) … The BDProbeTec™ Chlamydia trachomatis Amplified DNA Assay (BDPT) utilizes a strand displacement amplification (SDA) method coupled with a fluorescent energy transfer (ET) measurement in detecting the amplified product. Below are the links to the authors’ original submitted files for images. 1, Hannover, 30625, Germany, Rheumatologikum, Rathenau-Straße 13-14, Hannover, 30159, Germany, Division of Rheumatology, Rotes Kreuz Krankenhaus, St.-Pauli-Deich 24, Bremen, 28199, Germany, You can also search for this author in -80°C, or with nitric oxide can preserve the high detection rate. 1993, 38: 278-285. One ml of SF containing no added C. trachomatis EB was processed in each experiment as a negative control. J Clin Microbiol. The sensitivity and specificity compared to culture in detecting C. trachomatis in conjunctival specimens from infants with conjunctivitis were 93 and 96%, respectively. Kellogg ND, Baillargeon J, Lukefahr JL, Lawless K, Menard SW. J Pediatr Adolesc Gynecol. Laboratories employ different in-house methods for preparation of template DNA as well as different amplification systems. The resulting SF cell pellet was further processed by the different DNA extraction methods described below. This site needs JavaScript to work properly. Moreover, the existing in-house laboratory test systems have not yet been evaluated for their feasibility and sensitivity to detect C. trachomatis DNA in SF in clinical practice. The preferred method for chlamydia testing is the nucleic acid amplification test (NAAT) that detects the genetic material (DNA) of Chlamydia … statement and Each point on the graph indicates the detection limit of one serial dilution analysis. Numbers in boxes represent lowest reproducible detection limit of Chlamydia trachomatis EB/ml in synovial fluid. Antibiotic treatment for commensal organism growth in semen is not required. Positive Chlamydia cultures are confirmed for trachomatis by Chlamydia trachomatis by Transcription-Mediated Amplification (TMA) at no additional charge. The prevalence found represents a decrease from 10 to 2.9% of culture verified chlamydia during four years in comparable populations. Harms of screening asymptomatic men and nonpregnant women, including adolescents: 25 studies of tests for gonorrhea and 33 for chlamydia reported diagnostic accuracy. Sex transm dis. 153 million people globally have trichomoniasis, a higher prevalence than chlamydia and gonorrhea infections combined. Found inside – Page 89of therapy ( estimate 90 % cured ) , the high prevalence of infection , and the relatively low sensitivity of culture . Infection is less likely to persist ... The sensitivity of tissue culture ranges from 65 to 80 %. Part of Most people who are infected have no symptoms. Because PCR 1 allowed the highest sensitivity with several DNA extraction systems in contrast to the other PCR systems evaluated, in further analysis we restricted comparative detection of different sample preparation procedures based on the results of PCR 1. Bookshelf In infants with suspected chlamydial pneumonia, perform a nasopharyngeal swab for Chlamydia culture. Currently available rapid tests [ 38] are not approved for use on such specimens. In severe or complicated cases, consider sending bronchoalveolar lavage fluid for chlamydial culture as well. 1997, 36: 198-202. 1990, 28: 1968-1973. HIV, as per the recommendations in the HIV Screening and Testing Guide. EB were purified in a discontinuous gradient of Urografin® (Schering, Berlin, Germany) by ultracentrifugation, as described by Schmitz and colleagues [19]. Then, 140 μl chloroform (Baker, Deventer, the Netherlands) was added and samples were mixed for at least 30 seconds and subsequently centrifuged at 16,000 g for four minutes at room temperature. Clipboard, Search History, and several other advanced features are temporarily unavailable. in vitro to evaluate the sensitivity of different molecular biology methods and assays. A value of P ≤ 0.05 was considered significant in all such analyses. 2016 Aug 5;4(3):25. doi: 10.3390/microorganisms4030025. 10.1002/art.1780380718. Figure 1 summarizes the above described algorithm of sample analysis. Lower, but still reasonable, sensitivities to detect C. trachomatis EB DNA were achieved using alkaline lysis, QIAmp Tissue Kit® and QIAmp Stool Kit® in combination with the same amplification system. 0000008904 00000 n PMC This diversity most likely contributes to the variability of positive testing for C. trachomatis in clinical SF samples. 10.1002/art.1780310801. Sensitivity of Chlamydia-directed PCR and LCR testing for C. trachomatisEB DNA as a function of template preparation. 1996, 27: 1025-1034. %PDF-1.5 %���� Cervicitis in females. Anyone you share the following link with will be able to read this content: Sorry, a shareable link is not currently available for this article. DNA prepared by Qiaex II Gel Extraction Kit® + CTAB and DNA prepared by the Qiagen tissue kit® allowed detection of 10 C. trachomatis PBMO/ml SF in combination with number 1 PCR system. Found inside – Page 53Chlamydia sp. should be suspected in a person with a unilateral, chronic conjunctivitis and swabs should be taken for chlamydia culture/PCR or polymerase ... Clearly, the absence of an accurate gold standard is a difficulty for those evaluating newer diagnostic tests. Curr Opin Rheumatol. Ann Fam Med . 10.1093/rheumatology/40.8.920. Bacterial culture is simply a fancy way of saying "growing bacteria in a controlled setting." Fredlund H, Falk L, Jurstrand M, Unemo M. APMIS. Samples were considered positive when both duplicates were detected to be positive in the subsequent PCR analysis. 0000067720 00000 n PubMed  Genital infections are spread venereally, and respiratory infections usually by inhalation. 2020 Oct 8;2020:2892734. doi: 10.1155/2020/2892734. Conclusions: Workowski KA, Bachmann LH, Chan PA, Johnston CM, Muzny CA, Park I, Reno H, Zenilman JM, Bolan GA. MMWR Recomm Rep. 2021 Jul 23;70(4):1-187. doi: 10.15585/mmwr.rr7004a1. Although highly sensitive nucleic acid amplification tests (NAATs) are used to routinely diagnose chlamydial infection, C. trachomatis isolation by cell culture is still preferred for legal cases and epidemiological studies because of its high specificity; however, the sensitivity … https://www.verywellhealth.com/urine-testing-for-gonorrhea-and- On average, 0.1% (mean 0.0967%, standard deviation (SD) 0.0037) of monocytes was infected in each preparation analyzed. Primer set number 3 (Table 1) targets a sequence within the plasmid genome of C. trachomatis and conditions were used as first described by Wilkinson and colleagues [23]. Five different DNA-extraction methods were tested: 1) Alkaline lysis, 2) QIAex II Gel Extraction Kit®+ CTAB, 3) Chelex®-extraction, 4) QIAmp Tissue Kit® and 5) QIAmp DNA Stool Kit®. Schumacher HR, Magge S, Cherian PV, Sleckman J, Rothfuss S, Clayburne G, Sieck M: Light and electron microscopic studies on the synovial membrane in Reiter's syndrome. For chlamydia testing in men, the pooled sensitivities and specificities of each of the 3 assays Initially, immunofluorescence studies and RNA hybridization of synovial specimens were the first methods demonstrating intra-articularly persisting Chlamydia trachomatis [4, 5]. elementary bodies (EB) and human peripheral blood monocytes (PBMo) persistently infected with C. tr. Priem S, Rittig MG, Kamradt T, Burmester GR, Krause A: An optimized PCR leads to rapid and highly sensitive detection of Borrelia burgdorferi in patients with Lyme borreliosis. JF was responsible for organizational aspects of the study, collection of clinical samples, culture of C. trachomatis and performed DNA extraction, PCR analysis and drafted the manuscript. Uses of Culture and Sensitivity Urethral Swab Test. Clin Infect Dis. Diagnosis of sexually transmitted infections in children suspected of sexual abuse is challenging due to the medico-legal implications of test results. —Neither cell culture nor a rapid diagnostic test performed well for ensuring therapy of women with Chlamydia infections. Altogether, alkaline lysis and Qiaex II gel extraction kit® + CTAB gave reproducibly the highest detection rates in the C. trachomatis EB as well as in the C. trachomatis PBMO serial dilution analysis. Arthritis Rheum. Chlamydia culture, a cumbersome and time-consuming technique, was previously a prerequisite for obtaining DNA for WGS, but the introduction of culture-independent methodologies has resolved these glitches making WGS popular as the ultimate typing tool [9, 10, 99]. Epub 2009 Apr 15. Nucleic acid amplification tests in the diagnosis of chlamydial and gonococcal infections of the oropharynx and rectum in men who have sex with men. -EB/ml SF, 1 C. tr.-PBMo/ml, respectively, allowed most sensitive detection of the organism in combination with the C. tr.- omp1-(152 bp) PCR. 10.1093/rheumatology/36.2.198. The specificity is 99.5–100% 7, 21. Subsequently, neutralization was achieved by adding 12 μl 1 M Tris-HCl (pH 7.0). The two systems best suited for detection of C. trachomatis was applied to clinical samples of SF (data submitted elsewhere). Henning Zeidler and Jens G Kuipers contributed equally to this work. Köhler L, Nettelnbreker E, Hudson A, Ott N, Gérard H, Branigan P, Schumacher HR, Drommer W, Zeidler H: Ultrastructual and molecular analysis of the persistence of Chlamydia trachomatis (serovar K) in human monocytes. 10.1097/01.bor.0000126150.04251.f9. Hammer M, Nettelnbreker E, Hopf S, Schmitz E, Pörschke K, Zeidler H: Chlamydial rRNA in the joint of patients with Chlamydia induced arthritis and indifferentiated arthritis. Ann Rheum Dis. Biomed Res Int. Arthritis Res Ther 11, R175 (2009). Kent CK, Chaw JK, Wong W, et al. The patient's name and the source of the specimen must be on both the requisition and the specimen. (2) Given that 3 out of 4 infected women and 1 out of 2 infected men will be asymptomatic initially, the actual prevalence of disease is thought to … Detection limits of alkaline lysis dropped from an initial detection of 0.1 C. trachomatis EB/ml SF 100 fold to 10 detected C. trachomatis EB/ml SF and from 0.1 C. trachomatis PBMO/ml SF in immediate analysis to a 100-fold decreased detection rate to 1000 C. trachomatis PBMO/ml SF for stored samples (Table 4). 0000002256 00000 n 10.1136/ard.60.2.140. 2004, 16: 380-392. All other detection systems achieved equal or lower sensitivities in combination with the five DNA extraction methods investigated (Table 2). EB in known numbers were added to 1 ml SF in 10-fold decreasing numbers ranging from 103 to 10-3 C. trachomatis EB/ml SF. Method 4 and 5 differ in the contents of the added extraction buffers. 2004, 31: 499-507. Please enable it to take advantage of the complete set of features! CAS  We also tested four different PCR systems and one commercially available ligase chain reaction (LCR) protocol in use for urogenital samples in order to determine the most sensitive system to detect chlamydial DNA from SF. Primer system number 2 targets (Table 1) a different sequence in the C. trachomatis omp1 gene and was developed by Gérard and colleagues [9], and the conditions were described in several papers [4, 5, 9, 17]. In women with positive cervical culture results, the Gram stain results from the endocervix are 50-60% sensitive and 82-97% specific. Eight participants with CT-positive and 4 with NG-positive NAATs had negative culture results (P = 0.018 for CT urine NAATs vs. culture). Google Scholar. Branigan PJ, Gerard HC, Hudson AP, Schumacher HR, Pando J: Comparison of synovial tissue and synovial fluid as the source of nucleic acids for detection of Chlamydia trachomatis by polymerase chain reaction. 2002, 20: 185-192. Highest sensitivity (0.1 C. trachomatis EB/ml SF) was achieved with the Qiaex II Gel Extraction Kit® + CTAB followed by alkaline lysis, Qiagen Tissue Kit® and QIAmp DNA Stool Kit®, which detected repeatedly 1 C. trachomatis EB/ml SF in combination with PCR 1. Chlamydia, or more specifically a chlamydia infection, is a sexually transmitted infection caused by the bacterium Chlamydia trachomatis. People being evaluated or treated for a chlamydia infection should be screened for: Gonorrhea. Algorithm of sample analysis. The intracellular life style of chlamydia and the ability to cause persistent infections with low-grade replication requires tests with high analytical sensitivity to directly detect C. trachomatis (CT) in medical samples. The sensitivity and specificity data quoted in Table 1 are based on the new expanded gold LCX® analysis was performed in an LCR thermal cycler; patent of (Abbott, Abbott Park, IL, USA). Found inside – Page 338The sensitivity of these methods in general is comparable to the ... TREATMENT Chlamydial inclusion conjunctivitis is treated in neonates (0–7 days) with 20 ... 31, 32 We assume that the sensitivity of the gold standard test is 90% for C … J Rheumatol. Chelex® did not achieve sufficient sensitivity in C. trachomatis EB serial dilutions and was therefore not performed in the C. trachomatis PBMO assays. Wilkinson N, Kingsley G, Jones H, Sieper J, Braun J, Ward M: Detection of DNA from a range of bacterial species in the joints of patients with a variety of arthitides using a nested broad- range polymerase chain reaction. This advance has helped to improve the diagnosis and prevention of the major public health problem that is chlamydia infection. The Qiaex principle is based on the adsorption of DNA to silica gel particles in high salt. doi: 10.1128/JCM.01433-19. 0000005189 00000 n Article  2001, 40: 920-927. The product identity of PCR 1 was confirmed by hybridization using the digoxigenin hybridization protocol from Boehringer (Ingelheim, Germany) in combination with Dyna Beads (Dynal, Hamburg, Germany) for all analyzed samples. For the first time there are diagnostic tests for Chlamydia trachomatis that are more sensitive than tissue culture. Nucleic acid amplification tests (NAATs) are the most sensitive assays with a specificity similar to cell culture and are considered the method of choice for CT detection. Found insideIn this completely revised edition of the Clinical Virology Manual, Editor in Chief, Michael Loeffelholz, along with Section Editors, Richard Hodinka, Benjamin Pinsky, and Stephen Young, have complied expert perspectives of a renowned team ... In previous studies we already investigated the sensitivity of the Qiaex II gel extraction kit® in combination with the C. trachomatis omp1 152 bp PCR and have demonstrated that the DNA extraction method prior to PCR analysis influences the sensitivity to detect C. trachomatis DNA in synovial tissue [9] as well as in SF [17]. 0000012147 00000 n Briefly, aliquots of purified C. trachomatis EB were thawed and diluted to 20, 30, 40, 60, and 80 IFU/μl. Although culture is the legal standard, it is not the gold standard for the detection of C trachomatis . 1998, 25: 734-742. Introduction In the UK, practices for Chlamydia trachomatis (chlamydia) and Neisseria gonorrhoea (gonorrhoea) testing are not currently standardised. Julia Freise. Careers. Visualization of amplification products was performed by 2% agarose gel electrophoresis and ethidium bromide staining under ultraviolet light. 4. Would you like email updates of new search results? The most sensitive detection of chlamydial DNA was performed by DNA extraction by alkaline lysis which repeatedly detected 0.1 C. trachomatis PBMO/ml SF. To enhance the sensitivity of McCoy cells to C. trachomatis growth, cycloheximide was added to culture medium for cultivating infected cells. Arthritis Rheum. Moreover, some monocytes were observed to be infected with more than one C. trachomatis (data not shown). The sensitivity to detect C. tr.- DNA from SF is highly dependent on the DNA extraction method and the detection system applied. Informed written consent of each patient was obtained before storage of SF. Outside microorganisms may contaminate a specimen during each phase of handling. 8600 Rockville Pike At 10 days post infection, the cells were harvested and serially diluted in 10-fold decreasing steps in C. trachomatis-negative SF in a concentration ranging from 103 to 10-3 C. trachomatis PBMO/ml SF. 2005; 41(1):67-74. 10.1007/s004380050538. Comparison of nucleic acid amplification tests and culture techniques in the detection of Neisseria gonorrhoeae and Chlamydia trachomatis in victims of suspected child sexual abuse. Chlamydia is one of the most common bacterial sexually transmitted diseases (STD) in the United States and can cause serious complications if not treated. Psittacosis is acquired from infected birds. On average, six particles corresponded to 1 IFU in each dilution (slope = 6, r2 = 0.45; P = 0.0001). Found insideSince ocular infections are one of the most frequent occurrences in ophthalmology, the treatment for these infections must be fast, precise and effective. Bacterial culture is simply a fancy way of saying "growing bacteria in a controlled setting." Google Scholar. PubMed  He specializes in male infertility. 10.1016/0140-6736(92)90399-N. Kuipers JG, Andresen J, Kohler L, Schnarr S, Putschky N, Zeidler H, Wollenhaupt J: Evaluation of amplicor chlamydia PCR and LCX chlamydia LCR to detect Chlamydia trachomatis in synovial fluid. PCR and cytospin of the culture transport medium with chlamydia direct fluorescent antibody staining were used to adjudicate chlamydia culture-negative results. Serovar K was chosen because it causes urogenital tract infection and has been shown to cause ReA. C. trachomatis elementary bodies (EB) (serovar K) were cultured in Hep-2 cells as previously described [19]. Collect Charcoal ECS for culture and sensitivity – apply to slide, then place into charcoal medium.1 AND Obtain a 20ml first void urine for 1995, 38: 1005-1013. PCR 1 gave constantly most sensitive detection of C. trachomatis EB DNA in combination with all DNA extraction methods applied. These SF samples spiked with infected PBMO were processed with each of the above listed DNA extraction methods as in the EB studies. Kuipers JG, Nietfeld L, Dreses-Werringloer U, Koehler L, Wollenhaupt J, Zeidler H: Optimized sample preparation of synovial fluid for detection of Chlamydia trachomatis DNA by polymerase chain reaction. Moreover, PCR should overcome the limitations of clinical, urogenital, and serologic diagnosis of this form of ReA [16]. This might be due to the fact that intracellular persisting Chlamydia are showing an aberrant gene expression profile [27], which may influence the ease with which DNA extraction methods can release chlamydial DNA. <<669BE5C44FB93D4796E7CCB6F80E3C47>]/Prev 140826/XRefStm 1216>> Polymerase chain reaction (PCR) and ligase chain reaction (LCR) are used in research for detection of Chlamydia trachomatis (C. (1) Culture of rectal or pharyngeal specimens using a Chlamydia trachomatis-major outer membrane protein-specific stain. The DNA extraction methods alkaline lysis and Qiaex II Gel Extraction Kit® + CTAB allowed almost equal sensitivity limits according to our definition of sensitivity (lowest reproducible detection limit). Relative measures of test accuracy are less prone to bias in incomplete study … •Culture for chlamydia insensitive. Br J Rheumatol. in synovial fluid (SF). Jensen JS, Björnelius E, Dohn B, Lidbrink P: Comparison of first void urin and urogenital swab specimens for detetcion of Mycoplasma genitalium and Chlamydia trachomatis by polymerase chain reaction in patients attending a sexually transmitted disease clinic. Arthritis Rheum. This book will significantly enhance our ability to detect sexually transmitted infections and to conduct research to further our understanding of sexually transmitted diseases. In SF samples spiked with C. tr.-EB and with C. tr.-PBMo, alkaline lysis, detecting 1 C. tr.-EB/ml SF, 0,1 C. tr.-PBMo/ml SF and QIAmp gel extraction kit®+ CTAB detecting 0,1 C. tr. 1988, 31: 937-946. The apparent 2.407o false-posi-tive rate actually reflected the greater sensitivity of LCR: All 28 of the apparent false-positives had either chlamydial antigens Chlamydia trachomatis and Neisseria gonorrhoeae - Sensitivity and Specificity Comparison of BD DNA Assay to Culture (February 2007) Public Health Laboratories Ontario . Risk factors include multiple partners and failure to use condoms. two key statistics used to assess the performance of a screening test Google Scholar. Indication: required for medicolegal cases only; Sensitivity: 60-80%; Specificity: 100%; Urinalysis Leukocyte Esterase test (men) High Test Sensitivity for Chlamydia. Cookies policy. Tables 3 & 4 below provide updated sensitivity and specificity information for the Hologic® Aptima Combo 2® Assay for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae at urogenital sites for males and females. These results suggest that NAATs on urine, with confirmation, are adequate for use as a new forensic standard for diagnosis of CT and NG in children suspected of sexual abuse. Author information: (1)Department of Dermatovenereology, Tianjin Medical University General Hospital, Tianjin, China. Arthritis Rheum. The Qiagen products used for evaluation were supplied by courtesy of the Qiagen Company. The aim of the present study was to define a standardized and optimized test system to evaluate clinical SF samples for C. trachomatis DNA in routine laboratory analysis. Found inside – Page 1398Chlamydia trachomatis, Chlamydophila pneumoniae, and C. psittaci cause ... and cervical swab specimens increases culture sensitivity by approximately 20%?23 ... New gene amplification methods have replaced previous techniques, and first void urine samples have acquired an established position in Chlamydia diagnostics in both men and women. 1997, 255: 637-643. To our knowledge, some laboratories performing PCR analysis for routine diagnostic procedures store the extracted DNA at -20°C [25]. Evaluation of self-collected glans and rectal swabs from men who have sex with men for detection of Chlamydia trachomatis and Neisseria gonorrhoeae by use of nucleic acid amplification tests. 0000001385 00000 n California Privacy Statement, Rheumatology. Found inside – Page 890standard for the diagnosis of chlamydia. Cycloheximide-treated McCoy cells are the most ... When done correctly, cell culture has a sensitivity of 80%. can be identified by the test and suitable antibiotic therapy can also be determined on the basis of this test. 0000000016 00000 n This can be either clinician-collected or self-collected as follows: Remove the swab from the container, wipe the swab around the vaginal entrance, then PCR 4 also detected equal C. trachomatis EB/ml SF in combination with alkaline lysis (1 C. trachomatis EB/ml SF). Primer system number 1 (Table 1) was first described by Bobo and colleagues [24] and targets the C. trachomatis major outer membrane protein (omp1) gene; all assays were performed using the conditions described by Kuipers and colleagues [17]. Article 

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