Basics of molecular biology and its applications: III. Polymerase chain reaction and in situ hybridization.

Immunohistochemistry (IHC) has been successfully used to detect protein products and infective, microbial or viral particles in cells. One of its limitations is its inability to detect these agents when they are present in minute quantities. In situ hybridization (ISH) has an advantage over immunohistochemistry in its ability to demonstrate specific messenger RNA (mRNA). ISH can be combined with IHC in the same section to detect gene products as well as the mRNA encoding them. The polymerase chain reaction (PCR) offers an even more sensitive method. Genes code for proteins which are the final synthetic products expressed in the cells. Increased expression of the genes is reflected by an increased synthesis of protein products and higher levels of expression of transcribed mRNA. This principle is utilized in the application of ISH and PCR for the diagnosis of disease. The techniques of ISH and PCR are potentially useful in any situation that requires the examination of RNA or DNA. They have been used in the study of infectious, malignant and genetic diseases. With recent modifications and refinements, these techniques have transcended from the level of research tools to use in routine diagnostic laboratories.

[1]  R. Erasmus,et al.  Basis of molecular biology and its applications: II. An outline of common laboratory techniques used in molecular biology. , 1996, Papua and New Guinea medical journal.

[2]  R. Erasmus,et al.  Basics of molecular biology and its applications: I. Molecular biology in medicine: basic concepts. , 1996, Papua and New Guinea medical journal.

[3]  M. Alpers,et al.  The use of the polymerase chain reaction for more sensitive detection of Plasmodium falciparum. , 1995, Papua and New Guinea medical journal.

[4]  M. Alpers,et al.  Plasmodium falciparum: extensive polymorphism in merozoite surface antigen 2 alleles in an area with endemic malaria in Papua New Guinea. , 1994, Experimental parasitology.

[5]  L. Tavul,et al.  Plasmodium falciparum: a rapid technique for genotyping the merozoite surface protein 2. , 1993, Experimental parasitology.

[6]  Y. Wataya,et al.  DNA diagnosis of falciparum malaria using a double PCR technique: a field trial in the Solomon Islands. , 1993, Molecular and biochemical parasitology.

[7]  S. Narayanan.  Overview of principles and current uses of DNA probes in clinical and laboratory medicine. , 1992, Annals of clinical and laboratory science.

[8]  M. Tanner,et al.  Defective anion transport activity of the abnormal band 3 in hereditary ovalocytic red blood cells , 1992, Nature.

[9]  Huifen. Chen,et al.  Amplification of dengue 2 virus ribonucleic acid sequence using the polymerase chain reaction. , 1992, The Southeast Asian journal of tropical medicine and public health.

[10]  P. Jarolim,et al.  Deletion in erythrocyte band 3 gene in malaria-resistant Southeast Asian ovalocytosis. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[11]  R. Garruto,et al.  Highly divergent molecular variants of human T-lymphotropic virus type I from isolated populations in Papua New Guinea and the Solomon Islands. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[12]  R. G. Wright,et al.  Comparative in situ hybridisation study of juvenile laryngeal papillomatosis in Papua New Guinea and Australia. , 1990, Journal of clinical pathology.

[13]  W. Quint,et al.  Nucleic Acid Probe Techniques for the Routine Surgical Pathology Laboratory , 1988 .

[14]  H. Battifora,et al.  Abnormalities of protooncogenes in non‐small cell lung cancer. Correlations with tumor type and clinical characteristics , 1987, Cancer.

[15]  Bartow Sa.  Oncogenes and surgical pathology. , 1987 .

[16]  J. Bergh,et al.  Increased expression of N-myc in human small cell lung cancer biopsies predicts lack of response to chemotherapy and poor prognosis. , 1987, American journal of clinical pathology.

[17]  W. McGuire,et al.  Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. , 1987, Science.

[18]  H. Sather,et al.  Clinical implications of oncogene activation in human neuroblastomas , 1986, Cancer.

[19]  N. Kiviat,et al.  Detection and localization of human papillomavirus DNA in human genital condylomas by in situ hybridization with biotinylated probes , 1985, Journal of medical virology.

[20]  W. Tourtellotte,et al.  [Patient with progressive multifocal leukoencephalopathy by in situ hybridization: comparison of in situ hybridization using isotopic and biotinylated probes]. , 1985, No to shinkei = Brain and nerve.

[21]  A. Haase,et al.  Asymmetric replication of hepatitis B virus DNA in human liver: demonstration of cytoplasmic minus-strand DNA by blot analyses and in situ hybridization. , 1984, Virology.

[22]  D. Ward,et al.  Widespread presence of histologically occult cytomegalovirus. , 1984, Human pathology.

[23]  C. Burrell,et al.  Correlation Between Liver Histology and Markers of Hepatitis B Virus Replication in Infected Patients: A Study by In Situ Hybridization , 1984, Hepatology.

[24]  D. Ward,et al.  Detection of viral genomes in cultured cells and paraffin-embedded tissue sections using biotin-labeled hybridization probes. , 1983, Virology.