Biochips in the Diagnostic of Emerging Infectious Zoonotic Diseases
Development of Diagnostic Microchip for Study of Epidemiological Situation and Detection of Natural Areas of Emerging Infectious Zoonotic Diseases in Kyrgyz Republic
Tech Area / Field
- MED-DID/Diagnostics & Devices/Medicine
- MED-DIS/Disease Surveillance/Medicine
- AGR-DIS/Disease Surveillance/Agriculture
3 Approved without Funding
National Center of Cardiology and Internal Medicine, Kyrgyzstan, Bishkek
- Engelhardt Institute of Molecular Biology, Russia, Moscow\nNational Academy of Sciences of Kyrgyzstan / Biotechnology Institution, Kyrgyzstan, Bishkek
- Centers for Disease Control and Prevention (CDC), USA, GA, Atlanta
- The aim of the project
The aim of this project is development of a fast simultaneous real-time PCR analysis based on the biochip technology to identify the causative agents of infectious zoonotic and emerging diseases (anthrax, brucellosis, plague, tularemia, botulism, CCHF (Crimean Congo hemorrhagic fever), tick-borne encephalitis, and diseases caused by orthopoxviruses including Variola major). Identification of natural reservoirs of the pathogenic agents of interest and study of the epidemiological situation in the Kyrgyz Republic by the biochip-based method are tasks to be also performed in the framework of the Project.
IMPORTANT NOTE: All investigations will be performed using VACCINE AVIRULENT STRAINS of the corresponding causative agents.
2. The current status
An acute problem of public health and agriculture of the Kyrgyz Republic is emerging infectious diseases (EID), such as anthrax, brucellosis, plague, tularemia and sheeppox. The epidemiological situation of EID has been gradually worsening after the collapse of the USSR along with the general degradation of the economic situation in the Kyrgyz Republic. For example, the abundance of brucellosis among the local population grows yearly, on average 800 new cases of brucellosis are detected per year in the Kyrgyz Republic. There is a similar situation in agriculture, where approximately 2000 new cases of brucellosis in sheep were registered in 1999, and 9000 new cases were registered in the first 6 months of 2000. There are more than 1500 epidemiological areas of anthrax in the Kyrgyz Republic, and new cases of anthrax are reported every year. The spores of Bacillus anthracis were used in the terrorist attacks in theUSA, and in the Kyrgyz Republic the spores of Bacillus anthracis could also be used as a biological weapon by the extremist organizations as they have easy access to the natural regions of spores spreading. In the Kyrgyz Republic there are areas where the plague circulates permanently among rodents (mice and rats) and Kyrgyzstan and Kazakhstan remain natural reservoirs of plague. In the Soviet times, there was a special government service for plague prevention which was called antiplague stations. However after the collapse of the USSR, these stations actually stopped functioning in the Kyrgyz Republic due to insufficient financing. As a consequence, new cases of plague have been periodically reported there. There are also many areas with several zoonotic infectious diseases in the Kyrgyz Republic. Taking into account that most of the population work in agriculture, and the zoonotic areas are situated within some densely populated regions of the Kyrgyz Republic, there is an evident high risk of cross contamination of life stock animals and humans. Therefore, there is an urgent need to create a map of these zoonotic areas with accurately determined borders, and indicated types of animal-carriers, quantity and line of local population.
Rapid and reliable identification of anthrax, brucellosis, plague, tularemia and sheeppox presents a problem for many, if not all, countries. There are many different methods to detect these infections, such as: microbiological, virological, serological methods (ELISA), immunoblotting, mass-spectrometry, PCR, RT-PCR, “monoplex” real-time PCR, and etc. Unfortunately, these methods are relatively expensive, and usually a number of different methods (microscopic examination, serological test, ELISA, mass-spectrometry, PCR) need to be applied in order to reliably identify the pathogenic agent. Moreover, the traditional methods do not provide complete diagnostics, and they require prolonged time for study – from a few hours to a few months. For example, the sensitivity of the microbiological method of brucellosis detection is 53 to 90% at acute brucellosis (Ariza et al., 1995, Colmenero J. et al. 1996, Yagupsky P et al., 1999) and gets noticeably lower in chronical courses of brucellosis. The currently attained sensitivity of the PCR-based detection of Brucella melitensis is more than 96% (Morata P. et al., 2003).
Development and use of the real-time PCR based microchip approach will allow to avoid many significant problems associated with the traditional methods in identification of anthrax, brucellosis, plague, tularemia, botulism, CCHF (Crimean Congo hemorrhagic fever), tick-borne encephalitis, and diseases caused by orthopoxviruses including Variola major. The biochip-based method will also allow scientists to perform detection of the causative agents in one test and at the same time. It will improve the effectiveness of diagnostics of the diseases in samples of soil and blood of life stock animals.
All investigations will be performed using VACCINE AVIRULENT STRAINS of the corresponding causative agents. In case of botulism and CCHF the genome DNAs will be used as emulators.
The pision of National Center of Cardiology and Internal Medicine - Institute of Molecular Biology and Internal Medicine (IMBM) has solid experience in the field of biochip analysis and its staff includes highly qualified specialists in emergency infections. The biochip-based procedures were used in IMBM since 2003 to identify mutations in the different Mycobacterium tuberculosis strains resistant to antibiotics.
Engelhardt Institute of Molecular Biology, Russ. Acad. Sci., is one of the world’s leaders in the development of the biochip technology. Since 1988, the Biochip Technology Center of Engelhardt Institute of Molecular Biology has been developing microarrays (or biochips) consisting of three-dimensional hydrogel pads with immobilized probes (oligonucleotides, proteins, cells, etc.). Two main techniques of analysis have been designed to be applied directly on the biochip, namely hybridization of target nucleic acids with specific oligonucleotide probes immobilized in gel pads (Mikhailovich et al., 2001) and direct on-chip PCR amplification of target sequences (Strizhkov et al., 2000). By now, a number of causative agents of infectious diseases have been successfully identified by the biochip-based analysis (tuberculosis, anthrax, smallpox, viral hepatitis, etc). At present, the EIMB team is working out another approach: real-time on-chip multiplex PCR.
The International Science and Technology Center (ISTC) is an intergovernmental organization connecting scientists from Kazakhstan, Armenia, Tajikistan, Kyrgyzstan, and Georgia with their peers and research organizations in the EU, Japan, Republic of Korea, Norway and the United States.
ISTC facilitates international science projects and assists the global scientific and business community to source and engage with CIS and Georgian institutes that develop or possess an excellence of scientific know-how.