New and improved diagnostics for detection of drug-resistant pulmonary tuberculosis

O’Grady, Justin; Maeurer, Markus; Mwaba, Peter; Kapata, Nathan; Bates, Matthew; Höelscher, Michael; Zumla, Alimuddin

doi:10.1097/mcp.0b013e3283452346

Cited by 64 publications

(62 citation statements)

References 40 publications

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“…In addition, more than 85% of isoniazid-resistant strains are associated with mutations in the gene katG and the promoter region of the gene inhA (18,19). The WHO has recommended the use of some of these rapid tests, including GenoType MTBDR (Hain, Germany) and GeneXpert (Cepheid Corp., USA), based on their good performance (20). These molecular tools provide clinical reports with shorter turnaround times than traditional DST and have fewer biosafety concerns than the latter.…”

mentioning

confidence: 99%

Multicenter Evaluation of Genechip for Detection of Multidrug-Resistant Mycobacterium tuberculosis

Pang

Xing

Zhang³

et al. 2013

J Clin Microbiol

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e Drug-resistant tuberculosis (TB), especially multidrug-resistant TB (MDR-TB), is still one of the most serious threats to TB control worldwide. Early diagnosis of MDR-TB is important for effectively blocking transmission and establishing an effective protocol for chemotherapy. Genechip is a rapid diagnostic method based on molecular biology that overcomes the poor biosafety, time consumption, and other drawbacks of traditional drug sensitivity testing (DST) that can detect MDR-TB. However, the Genechip approach has not been effectively evaluated, especially in limited-resource laboratories. In this study, we evaluated the performance of Genechip for MDR-TB in 1,814 patients in four prefectural or municipal laboratories and compared its performance with that of traditional DST. The results showed that the sensitivity and specificity of Genechip were 87.56% and 97.95% for rifampin resistance and 80.34% and 95.82% for isoniazid resistance, respectively. In addition, we found that the positive grade of the sputum smears influenced the judgment of results by Genechip. The test judged only 75% of the specimens of "scanty" positive grade. However, the positive grade of the specimens showed no influence on the accuracy of Genechip. Overall, the study suggests that, in limited-resource laboratories, Genechip showed high sensitivity and specificity for rifampin and isoniazid resistance, making it a more effective, rapid, safe, and cost-beneficial method worthy of broader use in limited-resource laboratories in China.

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mentioning

confidence: 99%

Multicenter Evaluation of Genechip for Detection of Multidrug-Resistant Mycobacterium tuberculosis

Pang

Xing

Zhang³

et al. 2013

J Clin Microbiol

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“…Molecular-based tests, however, can produce results in Ͻ1 day (5,(8)(9)(10)(11)(12). The WHO has endorsed line probe hybridization and molecular beacon assays for the detection of drug resistance in highincidence settings (13).…”

mentioning

confidence: 99%

MTBDR plus and MTBDR sl Assays: Absence of Wild-Type Probe Hybridization and Implications for Detection of Drug-Resistant Tuberculosis

et al. 2016

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e Accurate identification of drug-resistant Mycobacterium tuberculosis is imperative for effective treatment and subsequent reduction in disease transmission. Line probe assays rapidly detect mutations associated with resistance and wild-type sequences associated with susceptibility. Examination of molecular-level performance is necessary for improved assay result interpretation and for continued diagnostic development. Using data collected from a large, multisite diagnostic study, probe hybridization results from line probe assays, MTBDRplus and MTBDRsl, were compared to those of sequencing, and the diagnostic performance of each individual mutation and wild-type probe was assessed. Line probe assay results classified as resistant due to the absence of wild-type probe hybridization were compared to those of sequencing to determine if novel mutations were inhibiting wild-type probe hybridization. The contribution of absent wild-type probe hybridization to the detection of drug resistance was assessed via comparison to a phenotypic reference standard. In our study, mutation probes demonstrated significantly higher specificities than wild-type probes and wild-type probes demonstrated marginally higher sensitivities than mutation probes, an ideal combination for detecting the presence of resistance conferring mutations while yielding the fewest number of false-positive results. The absence of wild-type probe hybridization without mutation probe hybridization was determined to be primarily the result of failure of mutation probe hybridization and not the result of novel or rare mutations. Compared to phenotypic culture-based drug susceptibility testing, the absence of wild-type probe hybridization without mutation probe hybridization significantly contributed to the detection of phenotypic rifampin and fluoroquinolone resistance with negligible increases in false-positive results. While the number of incident cases of tuberculosis (TB) has been steadily declining over the past decade, the prevalence of drug-resistant disease threatens to reverse these declines (1, 2). The World Health Organization (WHO) estimated that, in 2014, 3.3% of new and 20% of previously treated TB cases were multidrug-resistant (MDR) or were resistant to isoniazid (INH) and rifampin (RIF) (3). Furthermore, 9.7% of these MDR cases were extensively drug resistant (XDR) or were resistant to at least one fluoroquinolone (FQ) in addition to a second-line injectable drug (SLID), amikacin (AMK), kanamycin (KAN), or capreomycin (CAP). Although detection of M/XDR-TB is imperative for effective disease management, less than half of all MDR-TB cases were detected in 2014 (2-4). This detection gap is due, in part, to the lack of low-cost, rapid, and accurate diagnostic technologies for M/XDR-TB (4-6).Conventional culture-based phenotypic drug susceptibility testing (DST) methods can take months to yield results (7). Molecular-based tests, however, can produce results in Ͻ1 day (5,(8)(9)(10)(11)(12). The WHO has endorsed line probe hybridization and mole...

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“…MTBC'nin laboratuvar tanısında yıllardan beri kullanılmakta olan mikroskopi ve kültür, düşük duyarlılık ve geç sonuç verme gibi sınırlamalara sahiptir. Tanıda yaşanan bu sorunların aşılabilmesi için son yıllarda direkt klinik örneklerdeki MTBC'nin DNA'sını çoğaltmaya yö-nelik olarak düzenlenmiş NAAT'leri geliştirilmiş-tir ve TB tanısında rutin kullanımda yer almaya başlamışlardır (4)(5)(6)(8)(9)(10)(11)(12).…”

Section: Discussionunclassified

Klinik Örneklerde Mycobacterium Tuberculosis Saptanmasında PCR-ELISA Yönteminin Değerlendirilmesi

Karakayali¹,

Özkütük²

2016

Kocatepe Tıp Dergisi

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ÖZAmaç: Tüberküloz (TB) dünyada en önemli ölüm nedenlerinden biridir. Mycobacterium tuberculosis komplex'in (MTBC) neden olduğu hastalığın erken tanısı TB'un kontrolünde önemlidir. Bu amaçla son yıllarda TB'un hızlı tanı-sında nükleik asit amplifikasyon test (NAAT) yöntemleri yaygın olarak kullanılmaya başlamıştır. Klinik hasta örne-ğinden direkt olarak MTBC saptamaya yönelik NAAT yön-temlerinden biri de "Polymerase Chain Reaction-Enzyme Immunoassay" (PCR-ELISA)'dır. Bu çalışmada, PCR-ELISA temelli ticari bir test olan ProDect MYCOBACTERIUM TU-BERCULOSIS (bcs Biotect S.p.A., Cagliari, İtalya) testinin akciğer ve akciğer dışı örneklerde MTBC saptamadaki performansının araştırılması amaçlanmıştır. Gereç ve Yöntem:Çalışmada tüberküloz kuşkulu hastalardan alınan 100 klinik örnek (76 akciğer, 24 akciğer dışı örnek) mikroskopi, kültür ve ProDect MYCOBACTERIUM TUBERCULOSIS PCR-ELISA test yöntemi ile incelenmiştir. Bu testte, amplifiye ürünün saptanması "DNA enzyme immunoassay" (DNA-ELISA) yöntemi ile mikroplaklarda gerçekleştirilir. DNA-ELISA yöntemi mikroplak kuyucuklarına kaplanmış, streptavidin-biotin bağlı, tek iplikçikli prob DNA ile amplifiye edilmiş DNA'ların hibridizasyonu esasına dayanır. Aranan DNA ile prob arasındaki bağlanma anti-DNA fare monoklonal antikor kullanılarak saptanır. Bulgular: Altın standart yöntem olan kültür ile birlikte değerlendirildiğinde, tüm örneklerden elde edilen sonuçlara göre PCR-ELISA testinin duyarlılığı % 68.2, özgül-lüğü % 94.1 olarak belirlenmiştir. Akciğer örneklerinde duyarlılık ve özgüllük sırası ile % 71.1 ve % 93.5, akciğer dışı örneklerde ise % 61.9 ve % 100 olarak saptanmıştır. Yayma pozitif akciğer örneklerinde ise testin duyarlılığı % 83.3 ve özgüllüğü % 100 olarak bulunmuştur. Sonuç:Tüberkülozun hızlı tanısında PCR-ELISA yöntemi-nin kısmen çok fazla ekipman gerektirmeyen bir yöntem olması nedeni ile, özellikle yayma pozitif akciğer örnek-leri için, mikobakteriyoloji laboratuvarlarında tüberküloz tanısında rutin bir NAAT olarak değerlendirilebileceği sonucuna varılmıştır.Anahtar Kelimeler: Mycobacterium tuberculosis, PCR-E-LISA, tüberküloz, tanı. ABSTRACTObjective: Tuberculosis (TB) is one of the major causes of death worldwide. In the controlling of Tuberculosis (TB), early diagnosis of disease caused by Mycobacterium tuberculosis complex (MTBC) is vital. Hence, nucleic acid amplification tests (NAAT) for rapid diagnosis of TB came to be used widely in recent years. PCR-ELISA is a method which based on nucleic acid amplification based methods that are used for the direct detection of MTBC in clinical samples. The aim of study, was to evaluate the performance of ProDect MYCOBACTERIUM TUBERCULO-SIS (bcs Biotect S.p.A., Cagliari, Italia), a commercial test based on PCR-ELISA for the detection of MTBC in pulmonary and extrapulmonary clinical samples.Material and Methods: 100 clinical samples (76 pulmonary and 24 extrapulmonary) from patients suspected of TB were tested by microscopy, culture and ProDect MY-COBACTERIUM TUBERCULOSIS PCR-ELISA methods. In this test, detection...

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New and improved diagnostics for detection of drug-resistant pulmonary tuberculosis

Cited by 64 publications

References 40 publications

Multicenter Evaluation of Genechip for Detection of Multidrug-Resistant Mycobacterium tuberculosis

Multicenter Evaluation of Genechip for Detection of Multidrug-Resistant Mycobacterium tuberculosis

MTBDR plus and MTBDR sl Assays: Absence of Wild-Type Probe Hybridization and Implications for Detection of Drug-Resistant Tuberculosis

Klinik Örneklerde Mycobacterium Tuberculosis Saptanmasında PCR-ELISA Yönteminin Değerlendirilmesi

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