Potential Application of Digitally Linked Tuberculosis Diagnostics for Real-Time Surveillance of Drug-Resistant Tuberculosis Transmission: Validation and Analysis of Test Results

Ng, Kamela Charmaine S.; Meehan, Conor J.; Torrea, Gabriela; Goeminne, Léonie; Diels, Maren; Rigouts, Leen; André, Emmanuel

doi:10.2196/medinform.9309

Cited by 12 publications

(28 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The LODs recorded for classic Xpert and Ultra were higher (less sensitive) than for the LPAs. As a foremost example, classic Xpert performed relatively poorly in detecting heteroresistant mixtures with mutation S450L, which is by far the globally most prevalent allele in RR-TB (9, 21). Nevertheless, the 20-40 % LOD we found for classic Xpert was lower (more sensitive) compared to previous studies which recorded 65-90% LOD (14, 15).…”

Section: Discussionmentioning

confidence: 99%

“…This information is deemed useful for conducting research on Ultra data and could be practical in the field if export of the melt peak temperatures through the LIS port becomes feasible in future software updates, together with other raw data (e.g.wild-type and Mut melt probes associated with RR mutation). Export to connectivity solutions such as DataToCare (Savics, Belgium), GXAlert (SystemOne, USA), or C360 (Cepheid, USA), with possible integration into e-Health patient charts beyond tuberculosis diagnostics, allows to transform this data into usable information for the National TB Programs using a combination of unique patient ID and geographical information (9, 28). This may not only greatly benefit the remote resolution of discordant results to improve patient management, but also aids to build more systematic data on the prevalence and impact of heteroresistance on a programmatic level.…”

Section: Discussionmentioning

confidence: 99%

“…We subjected mixtures of the RR-TB bacillary suspensions to classic Xpert and Ultra (9, 10), and thermolysates to LPA-Hain and LPA-Nipro following manufacturer’s instructions. We recorded the LODs and corresponding RDT probe reaction per mutation type, in comparison with values cross-validated by CFU counts and variant quantification with Deeplex-MycTB.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

How well do routine molecular diagnostics detect rifampicin heteroresistance inMycobacterium tuberculosis?

Supply

Cobelens

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

Phone: 21 +32(0)33455345; Fax number: +32(0)32476333 22 2 ABSTRACT 23Rifampicin heteroresistance -where rifampicin-resistant and -susceptible tuberculosis bacilli 24 co-exist -may result in failed standard TB treatment and potential spread of rifampicin-25 resistant strains. Detection of rifampicin heteroresistance in routine rapid diagnostic tests 26 (RDTs) allows for patients to receive prompt and effective multidrug-resistant-TB treatment, 27 and may improve rifampicin-resistant TB control. 28 The limit of detection (LOD) of rifampicin heteroresistance for phenotypic drug susceptibility 29 testing by the proportion method is 1%, yet is insufficiently documented for RDTs. We 30 therefore aimed to determine, for the four RDTs (XpertMTB/RIF, XpertMTB/RIF Ultra, 31 GenotypeMTBDRplusv2.0, and GenoscholarNTM+MDRTBII), the LOD per probe and 32 mutation, validated by colony-forming-unit-counting and targeted deep sequencing 33 (Deeplex-MycTB). 34 We selected one rifampicin-susceptible and four rifampicin-resistant strains, with mutation 35 D435V, H445D, H445Y, and S450L respectively, mixed them in various proportions in 36 triplicate, tested them with each RDT, and determined the LODs per mutation type. MycTB revealed concordant proportions of the minority resistant variants in the mixtures. 38 The Deeplex-MycTB-validated-LODs ranged from 20-80% for XpertMTB/RIF, 20-70% for 39 Xpert Ultra, 5-10% for GenotypeMTBDRplusv2.0, and 1-10% for GenoscholarNTM+MTBII for 40 the different mutations. 41 Deeplex-MycTB, GenotypeMTBDRplusv2.0, and GenoscholarNTM+MDRTBII, provide explicit 42 information on rifampicin heteroresistance for the most frequently detected mutations. 43

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

How well do routine molecular diagnostics detect rifampicin heteroresistance inMycobacterium tuberculosis?

Supply

Cobelens

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Our findings confirm the ability of Ultra to unambiguously identify a wide range of RRDR mutations. With the unprecedented roll-out of XpertMTB/RIF and associated connectivity solutions, such as DataToCare (Savics, Belgium) and GXAlert (SystemOne, USA) (2), Ultra results may be exploited to rule-out transmission between RR-TB patients in a specific setting, distinguish relapse from reinfection, and resolve discordance between an RR Ultra result and a low-level RS phenotypic result due to a disputed mutation. For such applications, it is key that ΔTm values are included in the exported results.…”

Section: Discussionmentioning

confidence: 99%

“…The deluge of data produced by XpertMTB/RIF (Cepheid) can help improve global rifampicin-resistant tuberculosis (RR-TB) control strategies through molecular epidemiological surveillance (1, 2). Recently, a new version of the test – Xpert Ultra (hereinafter called Ultra) was released (3).…”

Section: Introductionmentioning

confidence: 99%

Xpert Ultra can unambiguously identify specific rifampicin resistance-conferring mutations

Deun

Meehan

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

Phone: +32(0)33455345 21 Fax number: +32(0)32476333 22 23 24 2 Introduction 25The deluge of data produced by XpertMTB/RIF (Cepheid) can help improve global 26 rifampicin-resistant tuberculosis (RR-TB) control strategies through molecular 27 epidemiological surveillance (1, 2). Recently, a new version of the test -Xpert Ultra 28 (hereinafter called Ultra) was released (3). Determining the relationship between RR-29 conferring rpoB mutations, Ultra probes, and melting temperature shifts (∆Tm)the 30 difference between mutant and wildtype melting temperaturesallows Ultra results to 31 be utilized for rapid detection of RR-TB strains and related underlying rpoB mutations. 32 Methods 33To validate the usefulness of Ultra results for predicting specific mutations, we analyzed 34 10 RS-TB and 107 RR-TB strains from the Belgian Coordinated Collections of 35Microorganisms 36 in the Institute of Tropical Medicine, Antwerp, Belgium. These strains harbor 36 unique 37 RR-conferring mutations determined by rpoB sequencing. 38 Results 39Overall, 31/32 (97%) mutations inside the Rifampicin Resistance Determining Region 40 (RRDR) were correctly identified by Ultra. Of concern, mutation His445Arg gave a "RIF 41Resistance INDERTERMINATE" result among 3/4 strains tested while it was reported as 42 RR in the initial validation study (3). The silent mutation Thr444Thr was not reported as 43 RR (Figure 1). The RR-conferring mutations on codons 170, 250, 299, 482, and 491 44 situated outside the RRDR were not detected. 45The probe reactions observed were largely in agreement with previous results (3) albeit 46 we noted that mutations Met434Val, Met434Thr and those in codon 435 were captured 47 only by probe rpoB2; Ser450Leu and Ser450Trp were captured by both probes rpoB3 48 3 and rpoB4a, His445Arg was captured only by probe rpoB3; and Lys446Gln was 49 captured only by probe rpoB4. 50All mutations except those in codon 450 were associated with a negative ∆Tm (Figure 51 2). The combination of ∆Tm values with the capturing probes enabled to differentiate 52 mutations in codons 428, 430, 431, 432, 434, 435, 441, 445, 446, and 452, including 53 disputed mutations (4) ( Table 1). Mutation Asp435Tyr was unambiguously distinguished 54 from Asp435Val through probe rpoB2|∆Tm|, while mutations Ser441Gln and Ser441Leu 55 were discriminated from the rest by|∆Tm| values of probes rpoB2 and rpoB3. Mutations 56 His445Asp and His445Tyr were distinguished from disputed mutations His445Leu and 57 His445Asn through probe rpoB3 |∆Tm|. Ser450Leu was distinguished from Ser450Trp 58 by probe rpoB4A |∆Tm| except for one strain with an outlier rpoB4A Tm of 70.9°C in 59 contrast to the other 13 strains with rpoB4A Tm of 73.3-73.8°C. The indeterminate result 60 associated with His445Arg may be caused by its |∆Tm|=1.8°C compared with |∆Tm| 61 typically exceeding 2°C for other mutations. 62 Conclusions 63Our findings confirm the ability of Ultra to unambiguously identify a wide range of RRDR 64 mutations. With the unprecedented roll-out of XpertMTB/RIF and asso...

show abstract

Identification and Characterization of Mycobacterial Species Using Whole-Genome Sequences

Riojas

Frank

Greenfield

et al. 2021

Methods in Molecular Biology

View full text Add to dashboard Cite

Potential Application of Digitally Linked Tuberculosis Diagnostics for Real-Time Surveillance of Drug-Resistant Tuberculosis Transmission: Validation and Analysis of Test Results

Cited by 12 publications

References 25 publications

How well do routine molecular diagnostics detect rifampicin heteroresistance inMycobacterium tuberculosis?

How well do routine molecular diagnostics detect rifampicin heteroresistance inMycobacterium tuberculosis?

Xpert Ultra can unambiguously identify specific rifampicin resistance-conferring mutations

Identification and Characterization of Mycobacterial Species Using Whole-Genome Sequences

Contact Info

Product

Resources

About