In this paper, the application of molecular biology technology in the prevention and control of mosquito-borne infectious diseases in China in recent years is reviewed
To provide reference for the prevention and control of infection and mosquito-borne infectious diseases in public health emergencies.
Molecular biology technology; Insect vectors; infectious disease
Insect-borne diseases are a group of diseases spread by pathogens carried by arthropods.
1992 There are 535 species registered by the International Arbovirus Center, of which 128.
Species are pathogenic to humans [1]. Among the legally reported infectious diseases in China, insect-borne infectious diseases account for the majority.
13 species, mosquitoes can spread not only viral diseases, but also viral diseases.
Parasitic diseases. Most of these diseases belong to natural focus diseases, some of which
Regional and timeliness, low morbidity and high mortality, mainly spread through the media.
Control for prevention and control [2]. In recent years, with the research of molecular biology technology and
Development, more and more widely used in the medical field, and made great progress.
This paper reviews the application of molecular biology technology in the diagnosis and control of mosquito-borne infectious diseases in recent years.
The applications in these fields are summarized as follows.
1 common molecular biology techniques [3]
1. 1 nucleic acid molecular hybridization technology
Molecular hybridization of nucleic acids, using the nucleus
According to the principle of base complementation of acid molecules, under certain conditions, double-stranded molecules are unwound into two.
Single strand, renaturated with heterologous DNA or RNA (single strand), if it is heterologous.
If there are complementary base sequences in some areas between DNA or RNA, they will appear repeatedly.
Hybridized nucleic acid molecules can be formed during sexual intercourse. Both sides of hybridization are nucleic acid sequences to be detected.
And a probe. Nucleic acid probes can be radionuclides, biotin or other active substances.
Qualitative marking. According to their sources and properties, they can be divided into cDNA probes and genome probes.
Needle, oligonucleotide probe, RNA probe, etc.
Classification: According to the test object, it can be divided into southern hybridization and southern hybridization.
Northern hybridization; According to the method used, it can be divided into point hybridization,
Slit hybridization and colony in situ hybridization; According to environmental conditions: divided into liquids.
Phase hybridization and solid phase hybridization.
1.2 polymerase chain reaction
Using the DNA molecule to be amplified as a template, a pair of DNA molecules interact with the template respectively.
Complementary oligonucleotide fragments are used as primers, and under the action of DNA polymerase, according to
The mechanism of semi-conservative replication extends along the template chain until the new DNA fusion is completed
All right. By repeating this process, the target DNA fragment can be amplified.
At the same time, newly synthesized DNA fragments can also be used as templates to make DNA.
The amount of synthesis increased exponentially.
Various application modes of PCR: degenerate primers.
Pcr, nested primer pcr, multiplex PCR
Pcr), reverse pcr (reverse pcr or reverse pcr), asymmetric pcr.
(asymmetric pcr), labeled pcr (lp-pcr) and color pcr, plus end.
Detection was carried out by pcr, anchored pcr or fixed pcr, slide pcr and reverse transcription pcr.
Rna and quantitative pcr.
1 3 DNA chip
Gene chip is also called DNA chip or DNA microarray.
(DNA microarray). In-situ synthesis using photoconductive or microprinting.
A large number of probe molecules with specific sequences are densely and orderly fixed on the mobile phase.
On the carrier to be treated, then the labeled sample to be tested is added for multiplex hybridization.
Hybridization is to analyze the existence of target molecules through the intensity and distribution of hybridization signals.
Number and sequence, so as to obtain the genetic information of the tested sample. Function: with communication
Large-scale, parallel, miniaturization and automation, but in practice, its research
High cost; The method is not standardized enough; The supporting software is not perfect.
Application of molecular biology technology in diagnosis of insect-borne diseases
2. 1 malaria
Huang Bingcheng et al [4] used pBF2 DNA fragment, which was labeled and used as a probe.
Plasmodium falciparum was detected in various plasmodium DNA samples. Plasmodium gene chip
The research contents of the parasite also include the discovery of new genes of plasmodium [5] and the regulatory network of transcription factors.
Collateral [6], adaptation mechanism of plasmodium to human host [7], and comparative genomic heterogeneity of plasmodium.
Cross analysis [8], molecular mechanism of antigen variation of plasmodium falciparum [9] and plasmodium attack
The mechanism of killing red blood cells [10] and so on.
2.2 Filariasis
Huang Zhibiao et al. [1 1] used PCR to detect bancroftian filariasis in blood.
Filariasis and microfilaria bancrofti can be detected in 100 positive blood samples. use
The results of 540 blood samples from Bancroftian filariasis monitoring point were all negative, and the microscope
The blood test was negative, too. The routine filariasis detection is blood collection at night, which has a large amount of information.
The results show that [12], SsP/PCR amplification system can be used to detect patients with bancroftian filariasis.
Circulating DNA in blood samples can be used for periodic or nocturnal periodic filariasis.
The daytime blood test fundamentally changed the diagnosis, monitoring and treatment of filariasis.
Work style.
2.3 Dengue fever
Zheng Kui et al. [13] used multiplex PCR technology to quickly identify four serotypes.
Skin virus, and multiple PCR of dengue virus in the same reaction tube.
Typing and identification confirmed that the epidemic situation of dengue fever in Guangdong in 2004 was type I.
Dengue virus; It is also reported that the application of oligonucleotide chip technology can simultaneously confirm the flow.
Infection and dengue virus [14]. There will be hope in areas long plagued by this disease.
Through the perfection of this technology, effective treatment and protection can be obtained.