RT-qPCR investigation of post-mortem tissues during COVID-19.

In 2020, there were numerous cases in Kazakhstan with clinical symptoms of COVID-19 but negative PCR results in nasopharyngeal and oropharyngeal swabs. The diagnosis was confirmed clinically and by CT scans (computed tomography). The problem with such negative PCR results for SARS-CoV-2 infection confirmation still exists and indicates the need to confirm the diagnosis in the bronchoalveolar lavage in such cases. There is also a lack of information about confirmation of SARS-CoV-2 infection in deceased patients. In this study, various tissue materials, including lungs, bronchi, and trachea, were examined from eight patients who died, presumably from SARS-CoV-2 infection, between 2020 and 2022. Naso/oropharyngeal swabs taken from these patients in hospitals tested PCR negative for SARS-CoV-2. This study presents a modified RNA isolation method based on a comparison of the most used methods for RNA isolation in laboratories: QIAamp Viral RNA Mini Kit and TRIzol-based method. This modified nucleic acid extraction protocol can be used to confirm SARS-CoV-2 infection by RT-qPCR in the tissues of deceased patients in disputed cases. RT-qPCR with RNA of SARS-CoV-2 re-extracted with such method from post-mortem tissues that were stored at -80 °C for more than 32 months still demonstrated high-yielding positive results.

Mycobacterium tuberculosis RD-Rio Strain in Kazakhstan.

Mycobacterium tuberculosis RD-Rio strains are still rare in the former Soviet Union countries and Asia. We describe a strain in Kazakhstan that belongs to the RD-Rio secondary branch, which is endemic to northwest Russia and eastern Europe. Although RD-Rio strains are frequently multidrug resistant, this heterogeneous branch included only drug-susceptible isolates.

Monitoring of pathogenic Borrelia burgdorferi sensu lato in the Almaty oblast, Kazakhstan.

Lyme borreliosis (LB) is one of the most common vector-borne diseases transmitted by ticks. It is caused by the Borrelia burgdorferi sensu lato (s.l.) genospecies. The Almaty oblast of Kazakhstan is considered endemic for LB. Nevertheless, there are significant gaps in the tick surveillance for LB agents in the region. We evaluated B. burgdorferi s.l. prevalence in 413 ixodid ticks collected from bitten people and the seroprevalence of antibodies to B. burgdorferi s.l. in 589 residents of the Almaty oblast, Kazakhstan. All samples were collected between 2018 and 2020. Borrelia burgdorferi s.l. DNA was detected in 24 % (9/38) of I. persulcatus ticks that attacked humans in the city of Almaty and the Talgar and Karasay districts. Multilocus sequence typing identified two novel B. afzelii sequence types in I. persulcatus. The seroprevalence of IgG antibodies against B. burgdorferi s.l. in the analyzed population was detected to be 5.8 %. The obtained results confirm active circulation of B. burgdorferi s.l. in the Almaty oblast of Kazakhstan and raise concern regarding LB preventive measures in the region.

Tick-borne pathogens and their vectors in Kazakhstan - A review.

Tick-borne diseases (TBDs), in particular Crimean-Congo hemorrhagic fever (CCHF) and tick-borne encephalitis (TBE), have been found to be expanding beyond their traditional geographic locations in Kazakhstan, which led to increased awareness and concern against TBDs in recent years. The tick fauna of Kazakhstan includes more than 30 species of human-biting ixodid ticks, recognized as carriers of a number of dangerous infectious pathogens that cause CCHF, TBE, tularemia, tick-borne rickettsioses and Lyme borreliosis. This paper presents the current knowledge of the main vectors of tick-borne pathogens (TBP) occurring in Kazakhstan, as well as a review of recent studies of ticks performed with the use of molecular methods being currently developed and widely utilized by Kazakh epidemiologists.

Molecular snapshot of Mycobacterium tuberculosis population in Kazakhstan: a country-wide study.

Republic of Kazakhstan is among the 27 high multidrug-resistant tuberculosis (MDR-TB) burden countries in the world. Here, we analyzed the population structure and phylogeography of Mycobacterium tuberculosis in Kazakhstan and impact of the identified genotypes on spread of drug resistant strains. A total of 159 M. tuberculosis isolates from different regions of Kazakhstan were typed using 24-MIRU-VNTR and spoligotyping, and the profiles were compared to the MIRU-VNTRplus and SITVIT_WEB databases. Eight isolates with double VNTR alleles were excluded from further analysis that was performed on 151 isolates. They were assigned to 10 families, Beijing (n = 109) being the largest and dominated by a single clonal cluster 94-32 and derived profiles (n = 101). The other families were represented mainly by LAM (n = 17), Ural (n = 8), NEW-1 (n = 3) and a new cluster named KAZ-1 (n = 8). Beijing, LAM and Ural isolates were detected in all parts of the country while Iran-specific family NEW-1 was found only in southern Kazakhstan (P = 0.001). A reduced scheme of 10 most polymorphic VNTR loci provided a discrimination similar to that achieved by 15-MIRU scheme and may be recommended for rapid preliminary screening of the clinical isolates in Kazakhstan. Multi-drug resistance was significantly more prevalent among Beijing (64/109) and LAM (7/17) strains compared to strains of other families (1/25; P = 0.0006 and 0.01, respectively). High prevalence of the genetically closely related MDR strains of the Beijing genotype found in different regions of Kazakhstan highlights their crucial impact on the current TB epidemic in this country.