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Composition and method for stabilizing and maintaining the viability of hardy microorganisms

A composition and microorganism technology, which can be applied in the directions of biochemical equipment and methods, determination/inspection of microorganisms, preservation of microorganisms, etc., and can solve the problems of intermittent shedding, increased cost, and no longer feasible for cultivation.

Inactive Publication Date: 2017-02-22
DNA GENOTEK
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, decontamination is known to significantly impair the viability of mycobacteria (Burdz et al., 2003; Krasnow and Wayne, 1966), and thus treatment also reduces the sensitivity of detection by culture
Second, the congenital nature of the disease produces low copy number and only intermittent shedding of the organism
A third issue concerns the inherent physiological properties of the mycobacteria themselves, which include i) aggregation, clumps, and bundles; ii) surface tension induced by the waxy cell wall; iii) buoyancy (ranging from 0.79 to 1.07 with a low mean In 1) (Silverstolpe, 1948); (iv) slow growth; and iv) thick cell walls make M. tuberculosis difficult to lyse
However, this approach requires a significant change in sample handling, increases the cost of handling, and for the historically "gold" standard test culture, the MTB organism is no longer viable
Similarly, Dorman et al. (2010) used alcohol (50% ethanol by volume) to preserve induced sputum samples; and made MTB no longer viable for culture

Method used

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  • Composition and method for stabilizing and maintaining the viability of hardy microorganisms
  • Composition and method for stabilizing and maintaining the viability of hardy microorganisms
  • Composition and method for stabilizing and maintaining the viability of hardy microorganisms

Examples

Experimental program
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Effect test

Embodiment 1

[0070] Example 1: Ambient temperature stabilization of sputum spiked with Mycobacterium tuberculosis in sample transport chemistry qualitative

[0071] One of the main goals of global control of TB in humans is laboratory diagnosis of Mycobacterium tuberculosis, the causative agent of TB, followed by appropriate treatment. Difficulties involving the collection, transportation, and processing of sputum samples have become major problems in current global TB control efforts. Mycobacterium tuberculosis is present in sputum samples that are often contaminated with other fast-growing microbial communities. The rapid growth of certain clinically relevant smaller species at ambient temperatures can kill or outgrow medically important pathogens. Consequently, delays in the availability of expertise or infrastructure for sample transport to the laboratory or for processing are problematic.

[0072] In high-burden countries, sputum acid-fast bacilli (AFB) microscopy services are u...

example 2

[0134] Example 2: Sample transport chemical liquefaction of sputum on contact

[0135] Sputum from humans is used to diagnose tuberculosis and to detect infection in cystic fibrosis patients. Bacteria are not evenly distributed in such highly viscous samples, which poses problems with acid-fast stains typically used to identify mycobacteria. Strikingly, this technique for diagnosing tuberculosis has been found to have a sensitivity of only about 50%, ie there is an equal chance of detecting the presence of TB versus not detecting it when the patient actually has the disease. This lack of sensitivity is due to the inability of existing sample preparation protocols to produce samples with uniform distribution of bacteria such that there is an equal probability of obtaining mycobacteria for staining from any part of the sample. Therefore, liquefaction and homogenization of sputum samples are critical to ensure accurate and representative sputum cultures and molecular diagnosti...

example 3

[0152] Example 3: Sample Transport in Tuberculosis Diagnosis Using Smear Microscopy, Culture and Molecular Diagnostic Assays chemical compatibility

[0153] Globally, approximately 2 billion people are infected with the potentially highly infectious Mycobacterium tuberculosis ("MTB"). Nearly 9 million people develop active disease each year, and 2 million die from the disease. Given the infectious nature of MTB, rapid and accurate diagnosis is an important element of MTB treatment and disease control.

[0154] Four common first-line drugs for anti-TB therapy are isoniazid (INH), rifampicin (RIF), ethambutol (EMB), and pyrazinamide (PZA). However, MTB strains can become resistant to one or more drugs, making cure difficult to achieve. RIF resistance is most commonly seen in multi-drug resistant (MDR-TB) strains and is reported with greater than 95% frequency in such isolates (Morris et al., 1995). MDR-TB is defined as tuberculosis disease caused by bacterial strains resi...

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Abstract

The present application is to provide a composition and method for stabilizing and maintaining the viability of hardy microorganisms from sample collection to downstream analysis. In particular, there is a method for preserving viable hardy bacteria, such as Mycobacteria, Bacillus anthracis, or Clostridium difficile, in a biological sample, comprising contacting the biological sample with a stabilization composition, wherein the stabilization composition comprises a chelating agent, a denaturing, a salt and has a p H between about 6 and about 11.

Description

technical field [0001] This application is related to the field of sample collection and storage. More specifically, the present application relates to compositions and methods for maintaining the viability of hardy microorganisms from sample collection to analysis. Background technique [0002] Tuberculosis (TB) remains a major global health problem. Most new infections and deaths occur in developing countries. In 2012 alone, the World Health Organization (WHO) estimated that 8.6 million people developed TB and 1.3 million died from the disease, including 320,000 deaths among HIV-positive individuals (Global Tuberculosis Report 2013, WHO). The number of TB deaths is unacceptably large, considering that most are preventable. WHO estimates that about one-third of the world's population, or 2 billion people, are infected with tuberculosis and are therefore at risk of developing active disease. Strikingly, WHO estimates that only one third of the 3.6 million smear-positive ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/24C12N1/04C12N1/20C12Q1/00C12Q1/68
CPCC12Q1/00C12N1/04C12Q1/24C12N1/20
Inventor H·哈依姆·伯恩博伊姆卡珊德拉·凯利·奇利诺比塔皮·雷雅克·奥斯卡·埃弗拉德·奈尔斯奥勒·马尔滕·德布莱恩
Owner DNA GENOTEK