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Introduction to the tests that can be carried out in PCR laboratories

Introduction to the tests that can be carried out in PCR laboratories

  • Time of issue:2022-11-10
  • Views:

(Summary description)Driven by the policy and the epidemic, many medical institutions around the world have established independent and compliant PCR laboratories. In addition to neo-crown nucleic acid testing, what else can PCR laboratories do? The following is a compilation of the tests that can be carried out in PCR laboratories.

Introduction to the tests that can be carried out in PCR laboratories

(Summary description)Driven by the policy and the epidemic, many medical institutions around the world have established independent and compliant PCR laboratories. In addition to neo-crown nucleic acid testing, what else can PCR laboratories do? The following is a compilation of the tests that can be carried out in PCR laboratories.

  • Categories:Blogs
  • Author:AIVD
  • Origin:
  • Time of issue:2022-11-10 17:33
  • Views:
Information

The PCR lab is also called gene amplification lab. PCR is short for Polymerase Chain Reaction, a molecular biology technique used to amplify specific DNA fragments, which can be seen as special DNA replication in vitro in living organisms. The DNA gene tracking system allows for a rapid grasp of the viral content of a patient's body with nanometer-level accuracy.


The construction of PCR laboratories is characterized by high costs, high personnel quality requirements, high laboratory operation costs, and a low volume of samples for routine testing; therefore, many hospitals did not build them before the new crown epidemic.


Driven by the policy and the epidemic, many medical institutions around the world have established independent and compliant PCR laboratories. In addition to neo-crown nucleic acid testing, what else can PCR laboratories do? The following is a compilation of the tests that can be carried out in PCR laboratories.

 


I. Molecular tests for infectious diseases


1. Hepatitis B virus nucleic acid detection

The use of real-time fluorescence quantitative PCR to quantitatively detect HBV DNA, so as to make a rapid early diagnosis of HBV infection. It is used for the auxiliary diagnosis of HBV infection and monitoring the efficacy of drug therapy for hepatitis B patients.


2. Hepatitis B virus genotyping test

About 60% of domestic HBV genotypes are C genes, and about 30% are B genes. PCR combined with Taqman technology is applied to detect the nucleic acid fragment of HBV type B/C specific DNA. It can be combined with clinical manifestations and other laboratory testing indicators to evaluate the patient's condition.


3. Hepatitis C virus nucleic acid detection

Using real-time fluorescence quantitative PCR detection technology, the quantitative detection of HCV RNA can be achieved by the change in fluorescence signal. It is used to assist in the diagnosis of HCV infection and monitor the efficacy of drug therapy for hepatitis C patients.


4. Hepatitis C virus genotyping

PCR combined with Taqman technology is applied to detect the specific RNA nucleic acid fragments of type 1b and type 2a HCV. Genotyping of HCV can be performed on specimens, and the patient's condition can be evaluated in combination with clinical manifestations and other laboratory test indexes.


5. Quantitative detection of Mycobacterium tuberculosis DNA

PCR combined with Taqman technology is used for fluorescence detection of specific DNA nucleic acid fragments of TB to make a rapid early diagnosis of TB infection. It is used for the auxiliary diagnosis of TB infection and monitoring the efficacy of drug treatment.


6. EBV DNA quantitative detection

Apply PCR combined with Taqman technology to detect EBV DNA by fluorescent PCR using EBV-specific primer probes. It is used for the auxiliary diagnosis of EBV infection and the monitoring of the efficacy of drug therapy for patients with other infections.


7. Cytomegalovirus DNA quantitative assay

PCR combined with Taqman technology is used for fluorescence detection of CMV-specific DNA nucleic acid fragments to make a rapid early diagnosis of CMV infection. It is used to assist in the diagnosis of CMV infection and monitor the efficacy of drug therapy.


8. Herpes simplex virus DNA quantitative testing

Using PCR combined with Taqman technology, fluorescent PCR typing of HSV type 1 and 2 specific DNA nucleic acid fragments using HSV specific primer probes is performed to make a rapid early diagnosis of HSV infection. It is used as an aid in the diagnosis of HSV infection and for monitoring the efficacy of drug therapy.


9. Mycoplasma pneumoniae DNA quantitative testing

PCR combined with Taqman technology is applied to detect specific DNA nucleic acid fragments of MP and CP simultaneously by fluorescent PCR, thus making a rapid early diagnosis of MP and CP infections. It is used for the auxiliary diagnosis of MP and CP infections and the monitoring of the efficacy of drug treatment.


10. Gonococcal nucleic acid test

Gonococcal nucleic acid testing is divided into nucleic acid extraction and thermostatic amplification. It is suitable for qualitative detection of gonococcal RNA in male urethral swab specimens and female cervical swab specimens, and the test results can be used for the auxiliary diagnosis of gonococcal infection and provide molecular diagnostic basis for early diagnosis of STDs and primary screening of high-risk groups of STDs.


11. Chlamydia trachomatis nucleic acid test

Chlamydia trachomatis nucleic acid testing is divided into nucleic acid extraction and thermostatic amplification. The test results can be used for the auxiliary diagnosis of Chlamydia trachomatis infection, providing a molecular diagnostic basis for the early diagnosis of patients and the initial screening of people at high risk for STDs.


12. Ureaplasma urealyticum nucleic acid test

Ureaplasma urealyticum nucleic acid testing is divided into nucleic acid extraction and thermostatic amplification. It is suitable for qualitative detection of Ureaplasma urealyticum RNA in male urethral swab specimens and female cervical swab specimens, and the test results can be used to assist in the diagnosis of Ureaplasma urealyticum infection and provide a molecular diagnostic basis for the early diagnosis of STDs and primary screening of people at high risk for STDs.


13. Human papillomavirus nucleic acid test

PCR, combined with fluorescent probe in vitro amplification and detection technology, is used for the genotyping of different types of human papillomavirus genotypes. It can be used to diagnose HPV DNA in the surface cells of clinical warts, cervical cells, and cervical mucus specimens and can be used as a diagnostic basis for clinical suspicion of warts, determination of the cause of cervical cancer, early diagnosis, and monitoring of treatment effects.


14. Qualitative nucleic acid test for human immunodeficiency virus

One-step RT-PCR system combined with fluorescent probes for in vitro amplification and detection technology. diagnosis of early infection in infants <18 months of age; diagnosis of acute HIV-1 infection to determine HIV infection.


15. Quantitative detection of human immunodeficiency virus nucleic acid

Application of RT-PCR technology combined with fluorescent probe detection technology. can be used to detect the effect of antiviral drug therapy; monitor disease progression, diagnose acute HIV-1 infection, and determine HIV-1 infection.


16. Group B Streptococcus nucleic acid detection

The real-time quantitative PCR technique combined with the Taqman probe is used to detect GBS DNA in specimens to make a rapid early diagnosis of GBS infection.


GBS is a parthenogenic anaerobic Gram-positive streptococcus with a carrier rate of about 30% in normal women, and is a conditional pathogen.

 


II. Molecular tests for hereditary diseases


1. Y chromosome microdeletion detection

The indications for routine testing include: male infertility patients before choosing single sperm follicular plasma injection or in vitro fertilization to produce offspring; patients with non-obstructive azoospermia; patients with severe oligospermia; male infertility patients with unknown causes before medication; and azoospermia patients before testicular biopsy.


Recommended indications for the test include: patients with oligospermia; patients with male infertility with normal sperm density but an unknown cause; patients with male infertility with cryptorchidism and/or varicocele; and patients with unexplained habitual abortion.


2. Phenylalanine hydroxylase gene mutation detection

The mutation in the PAH gene was detected by PCR amplification and gene sequencing. The mutation of the PAH gene is analyzed and diagnosed, and the mutation of the causative gene or the polymorphic marker linked to the causative gene is identified through the study of the prevalent patients and their family lines, so that a direct or indirect diagnosis can be made to confirm the mutation and identify the carriers. It is possible to confirm the diagnosis of patients before the appearance of PKU symptoms and to take timely treatment measures.


3. Genetic deafness gene testing

We use probe-specific PCR technology to detect mutations in human peripheral blood at the alleles of deafness-related genes, which can be used as an aid to diagnose the cause of clinical non-syndromic deafness (hearing loss) and to detect carriers of deafness genes in non-hearing loss population.


4. Genetic drug-related deafness gene detection

It is used to detect mutations related to drug-related deafness in the human mitochondrial genome to assist in clinical diagnosis, screening, and medication guidance for high-risk groups, as well as in epidemiological investigation, prenatal screening, and neonatal birth screening.


5. Thalassemia gene test

We use human peripheral blood specimens to detect 3 types of deletion alpha-thalassemia, 2 types of mutation alpha-thalassemia, and 11 types of mutation beta-thalassemia, which are common in Chinese people. It can assist in the diagnosis of people carrying the thalassemia gene in the high prevalence area and screen both couples before marriage, pregnancy, or the prenatal period to prevent the birth of children with thalassemia major and improve the quality of the population.

 


Ⅲ. The next generation sequencing(NGS) test items


1. BRCA gene mutation detection

BRCA is an important oncogene, including BRCA1 and BRCA2. The BRCA1/2 gene is an important biomarker to assess the risk of breast cancer, ovarian cancer, and other related cancers, as well as a biomarker to influence the choice of an individualized treatment plan for patients.


BRCA mutation testing of the blood and/or tumor tissues of patients with related cancers will help to better determine prognosis, select targeted drugs, choose chemotherapy regimens, assess the risk of disease in relatives of patients with family history of the disease under appropriate conditions, and help physicians select more accurate treatment plans based on the genetic status of patients.


2. Personalized genetic testing for lung cancer

For patients with advanced lung cancer who wish to receive targeted therapy or understand the mechanism of drug resistance, targeted lung cancer NGS testing can be used to cover multiple genes related to targeted lung cancer therapy, which has important clinical guidance significance for accurate and stratified diagnosis of lung cancer and selection of targeted drugs.


3. Breast cancer gene mutation detection

Mutation testing of breast cancer patients' blood and/or tumor tissues will help to better determine prognosis, select targeted drugs, choose chemotherapy regimens, and evaluate the risk of disease in relatives of patients with family history under appropriate conditions, and help physicians select more accurate treatment plans according to patients' genetic status.


4. BRAF-V600E gene mutation detection

The predictive role of the B-RAF mutation is well established, and the BRAF-V600E mutation can cause some patients with the wild-type KRAS gene to be insensitive to treatment with EGFR-TK1 and endothelial growth factor receptor monoclonal antibodies, so detection of the B-RAF mutation in tumor patients can be used to guide the use of targeted drugs.


5. Solid tumor-related gene detection

Tumor gene sequences are enriched by the hybridization capture method, and then high-throughput sequencing is performed on a sequencing platform. By detecting tumor-related gene hotspot region information and analyzing the mutation characteristics of these genes, we can assist clinicians in selecting appropriate therapeutic drugs for cancer patients and formulating perfect individualized treatment plans according to individual patient differentiation and combined with pharmacogenomics-related information.

 


Ⅳ. Drug susceptibility related molecular test items


1. Mycobacterium tuberculosis drug resistance gene polymorphism detection

The asymmetric PCR amplification technology of a real-time quantitative PCR instrument is used to detect mutations of Mycobacterium tuberculosis genes that are resistant to rifampicin and isoniazid. The rate of drug resistance of Mycobacterium tuberculosis is increasing worldwide, which has a great impact on the efficacy of drugs and the epidemic of tuberculosis, and the detection of drug resistance gene polymorphism of Mycobacterium tuberculosis is important for the treatment of tuberculosis.


2. Vancomycin resistance gene detection

Real-time PCR technology was used for nucleic acid amplification and target sequence detection of vancomycin-resistant enterococci. It guides clinical adjustment of antibiotic treatment regimen; rapid screening to facilitate early detection of hospital-acquired pathogens infected and carriers and timely interventions to avoid the spread and outbreak of hospital-acquired infections.


3. Methicillin-resistant Staphylococcus aureus drug resistance gene detection

Real-time PCR technology is used for nucleic acid amplification and target-sequence detection of MRSA. It is used to guide clinical adjustment of antibiotic treatment regimen; early detection of infected and carriers of infectious agents in hospitals, and timely interventions to avoid the spread and outbreak of hospital-acquired infections.


It should be clear that the laboratory project is closely related to the actual clinical needs. How to utilize PCR laboratories in a large number of primary and secondary care institutions in the future is a question that laboratory managers need to consider.


In addition, any other tests that can be carried out in PCR laboratories that are not listed in this article are welcome to be discussed in the comment section.

 


 

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