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Molecular Diagnostics Laboratory: Introduction, List of Tests, Specimen,Testing Method, and Keynotes

Introduction of Molecular Diagnostics Laboratory

As we know, simple diseases are due to either metabolic disorders or infectious agents (bacteria, viruses, fungi, parasites, prions). Molecular Diagnostics is a powerful tool for the identification of both conditions at genetic levels (e.g. metabolic disorders like cancer, and anemia while infectious conditions like COVID-19 and AIDS). It is applied for the detection of genomic variants, aiming to facilitate detection, diagnosis, sub-classification, prognosis, and monitoring response to therapy. This laboratory is also useful for those microbes which are hard to cultivate (Chlamydia, Rickettsia, viruses), time taken, and even dead microbes detection in specimens. RT-PCR is the gold standard for COVID-19 diagnosis to find out the etiological agent, SARS-CoV-2. Similarly, GeneXpert is the confirmatory test for tuberculosis. If I’m not wrong, today is the molecular era since such laboratories are having a greater impact on the treatment and care of patients and thus ‘Molecular Diagnostics’ is the game-changer for medicine.

Fig. Molecular Laboratory-A step of preparing reagent for RNA extraction of SARS-CoV-2

List of Molecular Biology Tests, Specimen, and Testing Methods

S.NoName of TestsSpecimenTesting Method
1.Alpha Thalassemia Mutation Detection (α3.7, α4.2)EDTA blood (3 ml)GAP PCR
2.Alpha Thalassemia Mutation Detection (?3.7, ?4.2)EDTA blood (3 ml)Real-Time PCR
3.Beta- Thalassemia Mutation Detection (IVS1-5 G-C, 619 bp deletion, cd 8/9 + G, IVS1-1 G-T, 41/42 – TTCT)EDTA blood (5 ml)Real-Time PCR
4.BK Virus/John Cunningham virus (JCV) Quantitative Real-Time PCRwhole blood (10 ml)Real-Time PCR
5.BRAF (gene) V600E MutationTissue (1 cm / 1 Block)Real-Time PCR
6.SARS-CoV-2 PCRNasopharyngeal swabRT PCR
7.Cytomegalovirus (CMV) Viral LoadEDTA blood (5 ml)Real-Time PCR
8.Dengue RNA PCREDTA blood (3 ml)Real-Time PCR
9.Duchenne/Becker muscular dystrophyEDTA blood (5 ml)Real-Time PCR
10.Epidermal Growth Factor Receptor gene (EGFR) Mutation AnalysisFFPE Block (1 cm / 1 Block)RT PCR
11.EGFR Mutation Analysis (Liquid Biopsy)EDTA blood (5 ml)RT PCR
12.EML-4 ALK Translocation by RT PCRFFPE Block (one slide)RT PCR
13.Fragile X (FMR 1) Mutation Screen EDTA blood (5 ml)Real-Time PCR
14.Gilbert Syndrome Mutation AnalysisEDTA blood (3 ml)RT PCR
15.Glucose-6 Phosphate Dehydrogenase (G6PD) Gene MutationsEDTA blood (3 ml)Real-Time PCR
16.Hepatitis B Virus (HBV) QualitativeEDTA blood (3 ml)Real-Time PCR
17.HBV Viral Load Real-Time PCREDTA blood (3 ml)Real-Time PCR
18.Hepatitis C virus (HCV) RNA PCR, QUALITATIVEEDTA blood (3 ml)Real-Time PCR
19.HCV RNA QUANTITATIVEEDTA blood (3 ml)Flocculation
20.HFE (Homeostatic Iron Regulator) gene mutationsEDTA blood (3 ml)Spectrophotometry
21.Human leukocyte antigen B27(HLA B27) PCREDTA blood (3 ml)Real-Time PCR
22.Human papillomavirus (HPV) DNA Detector Swab (3 ml)Real-Time PCR
23.Herpes simplex virus (HSV) DNA PCRSerum ( 1ml)Real-Time PCR
24.HPV (High Risk) (HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68)Swab (1 ml)Flocculation
25.Huntington Disease Molecular AnalysisEDTA blood (10 ml)Real-Time PCR
26.Inflammatory bowel disease (IBD) Screening Panel (ASCA IgA, ASCA IgG, ANCA)Serum ( 2 ml)CMIA
27.JAK2 Exon 12 Mutation EDTA blood (5 ml)Real-Time PCR
28.JAK2 V617F Mutation Detection Exon 14EDTA blood (5 ml)Real-Time PCR
29.JC/BK Virus (Quantatative)EDTA blood (3 ml)Real-Time PCR
30Mycobacterium tuberculosis PCR (GeneXpert)Sputum/Urine/BALReal-Time PCR
31.PML(Promyelocytic leukemia)/RARA (retinoic acid receptor alpha) t(15:17), Qualitative EDTA blood (3 ml)Real-Time PCR
32.PML/RaRa t(15:17), Quantitative EDTA Blood (3 ml)Real-Time PCR
33.SMA (spinal muscular atrophy) Carrier Detection EDTA blood (5 ml)MLPA Fragment Analysis
34.SRY (sex-determining region of the Y chromosome) Gene DetectionEDTA blood (3 ml)RT PCR
35.Thalassemia; Alpha Thalassemia Muation Detection (?3.7, ?4.2, ?FIL, ?SEA, ?THAI ) PNDTSwab (5 ml)Real-Time PCR
36.Vericella Zoster Virus (VZV) PCREDTA blood (5 ml)Real-Time PCR
Table: List of Molecular Tests, Specimen Requirements, and Testing Methods

Keynotes on Molecular Diagnostics

  • Gap-PCR stands for Gap-polymerase chain reaction.
  • MLPA stands for multiplex ligation-dependent probe amplification.
  • RT-PCR stands for a Reverse transcription-polymerase chain reaction.
  • CMIA stands for Chemiluminescent Microparticle Immuno Assay.
  • BK virus is a brief of the name of the first patient, from whom the virus was isolated in 1971 and the full name is not written because of personal identification following ethical guidelines.
  • Echinoderm microtubule-associated protein-like 4 anaplastic lymphoma kinase (EML4-ALK) is a fusion gene.
  • pANCA stands for Perinuclear Anti-Neutrophil Cytoplasmic Antibody while ASCA stands for Anti-Saccharomyces cerevisiae Antibody.
Medical Lab Notes

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