Whole-genome sequencing maps rpoB mutations that predict rifabutin susceptibility in multidrug-resistant tuberculosis

Rifabutin查看药品详情, a lesser-known cousin of the frontline tuberculosis drug rifampicin查看药品详情, may hold renewed value as a treatment option for multidrug-resistant tuberculosis (搜索)搜索该疾病 (MDR-TB)—but only for patients whose infecting strains carry specific genetic signatures. That is the central finding of a new whole-genome sequencing study from Shenzhen, China, published in BMC Infectious Diseases, which maps in detail how mutations in the bacterial RNA polymerase gene rpoB (搜索)搜索该靶点 determine whether MDR-TB isolates remain vulnerable to rifabutin.

Multidrug-resistant tuberculosis (搜索)搜索该疾病, defined by resistance to at least isoniazid and rifampicin查看药品详情, remains one of the most formidable challenges in global infectious disease control. Rifampicin resistance alone effectively disqualifies standard short-course regimens and forces patients onto longer, more toxic, and more expensive therapies. Yet rifampicin resistance does not always mean cross-resistance to every drug in the rifamycin class. Rifabutin查看药品详情, which shares its target with rifampicin—the beta subunit of bacterial DNA-dependent RNA polymerase, encoded by the rpoB (搜索)搜索该靶点 gene—can retain activity against some rifampicin-resistant strains of Mycobacterium tuberculosis. The clinical problem has been that no reliable genetic markers existed to tell clinicians which resistant infections would still respond to rifabutin and which would not.

The research team, led by Jing Gui, Jinli Li, Feng Wang, and Chuangyue Hong of the Shenzhen Center for Chronic Disease Control (搜索)搜索该词条, analyzed 183 MDR-TB isolates collected between 2013 and 2019 from patients in Shenzhen. For each isolate, the researchers determined the rifabutin查看药品详情 minimum inhibitory concentration (MIC) using broth microdilution carried out according to Clinical and Laboratory Standards Institute guideline M24-A2, with susceptibility defined as an MIC below 0.5 micrograms per milliliter. In parallel, whole-genome sequencing revealed each isolate's rpoB (搜索)搜索该靶点 mutation profile, its phylogenetic lineage, and the presence of compensatory mutations—secondary genetic changes that can restore bacterial fitness after resistance-conferring mutations impose a cost.

The statistical framework was deliberately layered. The team used Kruskal-Wallis tests to compare log2-transformed rifabutin查看药品详情 MIC values across groups of isolates defined by their rpoB (搜索)搜索该靶点 mutation type, Fisher's exact tests for categorical comparisons of susceptibility proportions, and multivariable linear regression models to disentangle the independent effects of rpoB genotype, bacterial lineage, and compensatory mutations.

The answer was unambiguous. The type of rpoB (搜索)搜索该靶点 mutation emerged as the primary determinant of rifabutin查看药品详情 susceptibility, but the key lay in which amino acid was altered. Isolates carrying the D435V mutation—the substitution of valine for aspartic acid at position 435 of the RNA polymerase beta subunit—consistently preserved susceptibility to rifabutin, with 66.7 percent of such isolates (4 of 6, 95 percent confidence interval 22.3 to 95.7 percent) falling below the susceptibility threshold. At the opposite end of the spectrum, the two most common rifampicin查看药品详情-resistance mutations worldwide, S450L and H445Y, conferred high-level rifabutin resistance: only 26.4 percent and 25.0 percent of isolates carrying these mutations, respectively, remained susceptible. Isolates harboring multiple rpoB mutations showed an intermediate phenotype, with 48.8 percent susceptible.

These findings carry substantial mechanistic logic. The rpoB (搜索)搜索该靶点 mutations that confer rifampicin查看药品详情 resistance cluster in a short region of the gene known as the rifampicin resistance-determining region, where amino acid substitutions alter the geometry of the drug-binding pocket. S450L, the single most frequent rifampicin-resistance mutation globally, replaces a serine with a bulky leucine, distorting the binding site in a manner that disrupts both rifampicin and rifabutin查看药品详情. H445Y produces a similar effect through a different chemical route. D435V, by contrast, appears to alter the pocket enough to block rifampicin while leaving sufficient structural compatibility for rifabutin.

Perhaps the most clinically consequential finding concerned the two mutations that dominate the global rifampicin查看药品详情-resistance landscape. Because S450L and H445Y reliably predict high-level rifabutin查看药品详情 resistance, the study suggests that a positive molecular test for rifampicin resistance should not automatically be interpreted as rifabutin eligibility. Instead, the specific mutation matters enormously. A patient whose isolate carries D435V may still benefit from rifabutin-containing therapy, while a patient with S450L almost certainly will not. In settings where whole-genome sequencing is already deployed for tuberculosis diagnosis and surveillance, this information comes essentially free of charge—an added layer of therapeutic intelligence extracted from data already being generated.

The regression models added nuance beyond the rpoB (搜索)搜索该靶点 story. After adjusting for rpoB genotype, the researchers found that bacterial lineage exerted a modest but statistically significant independent effect on rifabutin查看药品详情 MIC. Isolates belonging to lineages other than Lineage 2—the so-called Beijing lineage, which dominates in East Asia—had lower rifabutin MICs, with an adjusted beta coefficient of −0.90 (95 percent confidence interval −1.77 to −0.04, p = 0.041). Expressed differently, Lineage 2 strains tended to show higher ratios of rifampicin查看药品详情 to rifabutin MIC values, hinting at lineage-specific differences in how the rifamycin-binding pocket tolerates each drug.

Compensatory mutations, by contrast, told a simpler story. These secondary changes, which arise to restore the transcriptional efficiency of a drug-resistant RNA polymerase, did not show any independent association with rifabutin查看药品详情 MIC after adjustment for rpoB (搜索)搜索该靶点 genotype (adjusted beta = −0.34, 95 percent confidence interval −1.02 to 0.33, p = 0.323). This null result indicates that compensatory evolution in rifampicin查看药品详情-resistant tuberculosis does not inadvertently alter rifabutin susceptibility, allowing clinicians and genomic surveillance systems to focus their predictive attention on the primary resistance mutations themselves.

The study also probed whether the level of rifabutin查看药品详情 resistance correlated with transmission dynamics, using molecular clustering of whole-genome sequences as a proxy for recent transmission. It did not: the odds ratio linking high-level resistance to clustering was 1.95 (95 percent confidence interval 0.84 to 4.80, p = 0.129), falling short of statistical significance. While the point estimate raises the possibility that highly resistant strains might transmit somewhat more readily, the data do not support a firm conclusion, and the authors treat this as exploratory.

The implications reach well beyond Shenzhen. Rifabutin查看药品详情 has long occupied an awkward position in tuberculosis therapeutics—chemically capable of activity against some rifampicin查看药品详情-resistant strains, but rarely used against MDR-TB because susceptibility could not be predicted. The demonstration that rpoB (搜索)搜索该靶点 genotype, particularly D435V, functions as a robust biomarker of rifabutin susceptibility opens the door to genotype-guided rifabutin prescribing within existing genomic surveillance infrastructure. As whole-genome sequencing becomes more affordable and more widespread in high-burden countries, the marginal cost of applying these findings approaches zero. The study also strengthens the case for building rifamycin cross-resistance prediction into international drug-resistance databases and treatment guidelines, where such nuance is currently absent.

Important caveats remain. The D435V group was small—only six isolates—which is reflected in the wide confidence interval around the susceptibility estimate, and the findings derive from a single Chinese city where Lineage 2 predominates. Validation in other geographic and lineage contexts will be needed before rpoB (搜索)搜索该靶点 genotype can formally guide rifabutin查看药品详情 use in clinical trials and treatment programs. Still, the study delivers what precision medicine for tuberculosis has lacked for this drug class: a clear, quantitatively grounded map of which resistance mutations preserve rifabutin activity and which destroy it.