Rv3737 is required for Mycobacterium tuberculosis growth in vitro and in vivo and correlates with bacterial load and disease severity in human tuberculosis.

BACKGROUND: Rv3737 is the sole homologue of multifunctional transporter ThrE in Mycobacterium tuberculosis (Mtb). In this study, we aimed to investigate whether this transporter participates in vitro and in vivo survival of Mtb. METHODS: To characterize the role of Rv3737, we constructed and characterized a Mtb H37RvΔRv3737. This strain was evaluated for altered growth rate and macrophage survival using a cell model of infection. In addition, the comparative analysis was conducted to determine the association between Rv3737 mRNA expression and disease severity in active pulmonary TB patients. RESULTS: The H37RvΔRv3737 strain exhibited significantly slow growth rate compared to H37Rv-WT strain in standard culture medium. Additionally, the survival rate of H37Rv-WT strain in macrophages was 2 folds higher than that of H37RvΔRv3737 at 72 h. A significantly higher level of TNF-α and IL-6 mRNA expression was observed in macrophages infected with H37RvΔRv3737 as compared to H37Rv-WT. Of note, Rv3737 expression was significantly increased in clinical Mtb isolates than H37Rv-WT. The relative expression level of Rv3737 was positively correlated with lung cavity number of TB patients. Similarly, the higher Rv3737 mRNA level resulted in lower C(t) value by Xpert MTB/RIF assay, demonstrating that a positive correlation between Rv3737 expression and bacterial load in TB patients. CONCLUSIONS: Our data takes the lead in demonstrate that the threonine transporter Rv3737 is required for in vitro growth and survival of bacteria inside macrophages. In addition, the expression level of Rv3737 may be associated with bacterial load and disease severity in pulmonary tuberculosis patients.

Successful Treatment of a Multidrug-Resistant Tuberculosis Patient with a Negative Xpert MTB/RIF Test for Rifampicin-Resistant Tuberculosis in Guizhou Province of China: A Case Report.

The Xpert MTB/RIF (Xpert) assay recommended by the World Health Organization (WHO) can be used to simultaneously detect Mycobacterium tuberculosis complex (MTBC) and rifampicin (RIF) resistance associated mutations. However, if Xpert testing results are negative for RIF resistance because mutations outside the RIF resistance-determining region (RRDR) are not detectable by the assay, patients with RIF-resistant/multidrug-resistant tuberculosis (RR/MDR-TB) will be treated inappropriately for several weeks prior to obtaining the drug susceptibility testing (DST) results. Here, we report a rare case of TB in Guizhou Province of China that was identified as RIF-susceptible by the Xpert MTB/RIF assay, but later was confirmed as MDR-TB by DST, and its successful treatment with effective second-line anti-TB drugs. We detected a rare rpoB mutation (Ile572Phe) in clinical samples of this patient, highlighting the importance of using other methods such as PCR and sequencing to complement the Xpert MTB/RIF assay for the routine diagnosis of RR/MDR-TB because of the limited scope of the assay. These complementary methods allow for the detection of rare rpoB mutations outside the RRDR and can be beneficial when used in geographical locations where such rpoB mutations are frequently reported. However, these methods may not be feasible for resource-limited settings.