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It is necessary to institute a specific therapy at the earliest in infectious diseases. An early specific method of diagnosis is essential for this purpose.PCR is a more sensitive and rapid method and is also specific for the infectious agent being tested for. The results help physicians and surgeons to institute the specific therapy required for the treatment of the disease at the earliest possible time.

The In house PCRs are applied on all kinds of clinical specimens including body fluids, biopsy specimens, pus for 14 infective agents from the year 2006 onwards at VRF Referral Laboratory. There are 120 hospitals in and around Chennai making use of the VRF Referral Laboratory for rapid detection of the genome of the infective agents associated with the disease.

Mycobacterium tuberculosis is an obligate, acid – fast, Gram – positive bacilli infecting human beings. Its growth is slow and needs 3-4 weeks for a visible growth. The organism causes “human type” of tuberculosis in man. Primary infection is usually accompanied by mild or no symptoms, but this infection might get complicated due to the spread of organism to kidney, spleen, bone marrow, central nervous system and other organs including bone. The standard conventional methods of diagnosis are time consuming, (4-6 weeks) and are not always successful, especially in cases of extra pulmonary tuberculosis. electricity transformer near house By application of the nested PCR, M. tuberculosis genome can be detected within a day which aids in initiating the appropriate therapy immediately.

Toxoplasmosis is an infectious disease affecting both animals and humans which is caused by the protozoan Toxoplasma gondii. It is generally asymptomatic. CNS and ocular Toxoplasmosis is known to occur specially in immunocompromised persons. In pregnant women however, the infection acquires a special significance as the parasite may enter the fetal circulation though the placenta and cause congenital toxoplasmosis.

Cytomegalovirus belongs to Herpesviridae family; is recognized as a major causative agent of various diseases which occur in all age groups including hepatitis, pneumonia, mononucleosis, acute retinal necrosis especially in immunocompromised persons such as HIV/AIDS patients, renal transplanted patients and patients on immunosuppressive therapy. These diseases can be fatal. CMV is also known to cause congenital abnormalities in infants infected during fetal stage.

Herpes simplex virus is a widespread human pathogen. Serologically distiniguishable serotypes HSV-1 and HSV-2 cause herpetic lesions. Both the viruses infect epithelial cells. HSV-1 is normally associated with oral infections and lesions above waist and HSV-2 is associated with genital infections and lesions below waist. HSV is well known to undergo latency and cause various clinical diseases when it gets activated under conditions like stress, fever, immunosupression etc. The virus also causes HSV encephalitis and other organ system diseases. Detection of the virus in the clinical specimen from the lesion is diagnostic. e 87 gasoline PCR has been shown to be a rapid, highly sensitive and specific method for detection of the virus.

Harishankar A, Jambulingam M, Gowrishankar R, Venkatachalam A, Vetrivel U, Ravichandran S, Yesupadam SM, Madhavan HN. Phylogenetic comparison of exonic US4, US7 and UL44 regions of clinical herpes simplex virus type 1 isolates showed lack of association between their anatomic sites of infection and genotypic/sub genotypic classification. Virol J. 2012 Mar 14; 9(1):65. [Epub ahead of print]

Adenovirus is a DNA virus. These viruses can undergo latent infection in lymphoid tissues. 51 serotypes classified into 6 subgenera (A- F). Adenovirus infections involve the respiratory gastrointestinal tracts and the eye. Adenovirus infections are very common, most are asymptomatic. Most people have been infected with at least 1 type by age 15. Virus can be isolated from the majority of tonsils/adenoids surgically removed, indicating latent infections. gas pump emoji It is not known how long the virus can persist in the body, or whether it is capable of reactivation after long periods, causing disease (It is hard to isolate this occult virus as it may be present in only a few cells) it is known that Adenovirus is reactivated during immunosuppression, e.g. in AIDS patients.

Chlamydia trachomatis belongs to the family chalmydiae and are primarily human pathogens. They are obligate intracellular parasites. Chlamydia trachomatis is known to cause urethritis, epididymitis, proctitis, cervicitis, pelvic inflammatory disease, infant pneumoniae and conjunctivitis and trachoma. A variety of causes can bring about the signs and symptoms of these diseases, so diagnosis is required for prompt and accurate treatment. v gashi 2012 However, chlamydial infections are often asymptomatic.

Varicella Zoaster virus (VZV) belongs to the herpes virus family. Varicella is an acute viral disease caused by the virus characterized with fever, malaise and a maculopapular rash that changes within hours of formation of vesicles that remain 3 to 4 days. Lesions appear in the scalp, face neck trunk, mucous membranes of the mouth and upper respiratory tract. In adults, the fever and general clinical status can be severe, but the disease is rarely fatal. Death occurs in adults due to primary viral pneumonia and in children, secondary bacterial infections or central nervous system involvement. immunocompromised person are at higher risk of a general dissemination of the disease with fatal outcome. Infection at the beginning of pregnancy rarely gives congenital malformations. Reactivation of VZV is usually manifested as zoster.

Real-time PCR has engendered wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility, and the reduced risk of carry-over contamination as it does not require any post PCR handling. The real-time PCR method has a very large dynamic range of starting target molecule determination (at least five orders of magnitude) and is extremely accurate and less labor-intensive than current quantitative PCR methods.

The assay significantly has higher reliability of the results compared with conventional PCR, because with real-time PCR, the whole amplification profile is known. Individual reactions deviating in their amplification efficiency (e.g. owing to the presence of polymerase inhibitors) can be identified easily. Quantitative real time PCR is more precise than end-point determinations. The assay is commonly recommended wherever the exact number of the target is required and also to monitor prognosis in patients on therapy.

Real-time PCR has gained wider acceptance of the PCR due to its improved rapidity, sensitivity, reproducibility, and the reduced risk of carry-over contamination as it does not require any post PCR handling. The real-time PCR method has a very large dynamic range of starting target molecule determination (at least five orders of magnitude) and is extremely accurate and less labor-intensive than current quantitative PCR methods.

The assay significantly has higher reliability of the results compared with conventional PCR, because with real-time PCR, the whole amplification profile is known. 4 main gases in the atmosphere Individual reactions deviating in their amplification efficiency (e.g. owing to the presence of polymerase inhibitors) can be identified easily. Quantitative real time PCR is more precise than end-point determinations. The assay is commonly recommended wherever the exact number of the target is required and also to monitor prognosis in patients on therapy.

Human immunodeficiency virus (HIV) is a lentivirus (a member of the retrovirus family) that can lead to acquired immunodeficiency syndrome (AIDS), a condition in humans in which the immune system begins to fail, leading to life-threatening opportunistic infections. Infection with HIV occurs by the transfer of blood, semen, vaginal fluid or breast milk. Within these body fluids, HIV is present as both free virus particles and virus within infected immune cells. The four major routes of transmission are unprotected sexual intercourse, contaminated needles, breast milk, and transmission from an infected mother to her baby at birth (Vertical transmission). The persistence of proviral human immunodeficiency virus type 1 (HIV-1) DNA reservoir represents one of the major drawbacks to the total eradication of HIV-1. The quantitative determination of proviral HIV-1 DNA load offers significant therapeutic information, especially when the HIV-1 RNA levels drop below the detectable limits during the highly active retroviral therapy (HAART) treatment. electricity lessons grade 6 Moreover, the detection of HIV-1 proviral DNA is an important diagnostic marker in the evaluation of HIV-1 infection

Chikungunya is a RNA virus belongs to the family Togaviridae, genus Alphavirus. The disease Chikungunya also known as Chikungunya virus disease or Chikungunya fever is characterized by severe, sometimes persistent, joint pain (arthritis), as well as fever and rash. The disease is spread by the bite of infected mosquitoes and the clinical symptom resembles that of dengue fever. It is rarely life-threatening. In India the disease is more prevalent in Andhra Pradesh, Karnataka, Maharasthra, Tamil Nadu, Madhya Pradesh, Gujarat, Kerala, A & N Island, GNCT of Delhi, Rajasthan, Pondicherry, Goa. The virus isolation technique is cumbersome, and serological diagnosis is so often not reliable. The best method of detection of the virus is real time PCR.

Cytomegalovirus is a DNA beta-herpes virus and is ubiquitous in nature. Infection caused by the virus is usually silent in immunocompetent individuals, although acute CMV infection may also cause a brief mononucleosis-like malaise in immunocompetent adults. It is proven to establish latency in macrophages. Following infection, the virus resides in endothelial cells, macrophages, or granulocyte stem cells and may cause re infection if the host is rendered immunosuppressed, as by HIV or by immunosuppressive agents used during transplantation and chemotherapy. From infected pregnant women the virus gets transmitted to fetuses causing congenital defects, as they have poorly developed immune systems. As normal population is known to harbor baseline copy number of virus, estimation of quantity of virus is very important to distinguish normal subjects from diseased especially in blood samples. electricity sources in us Real time PCR is the best method to quantify the virus copy numbers.

Hepatitis B virus is an hepadnavirus — hepa from hepatotrophic and dna because it is a DNA virus. The virus has a circular genome composed of partially double-stranded DNA. The viruses replicate through an RNA intermediate form by reverse transcription.Hepatitis B causes hepatitis an infection and inflammation of the liver. The disease may lead to carcinoma of liver if not intervened.Although serological markers are available for the detection of the viral antigens, DNA amplification techniques are preferred due to the marked sensitivity of the assay. Quantification by real time PCR is advised for monitoring viral load for the prognosis.

The Hepatitis C virus (HCV) is a small (50 nm in size), enveloped, single-stranded, positive sense RNA virus. HCV causes Hepatitis C, an infectious disease of the liver, The infection is often asymptomatic, but once established, chronic infection can progress to fibrosis, and cirrhosis . HCV is spread by blood-to-blood contact. Most people have few symptoms after the initial infection, yet the virus persists in the liver in about 80% of those infected. gas x dosage for dogs Persistent infection can be treated with medication, such as interferon and ribavirin, but only a minority is cured. The disease can be diagnosed by serological markers. The HCV viral load is an important factor in determining the probability of response to interferon-based therapy which is done by real time PCR.

Mycobacterium tuberculosis is an obligate, acid fast, Gram positive bacilli infecting human beings. Its growth is slow and needs 3-4 weeks for a visible growth. Primary infection is usually accompanied by mild or no symptoms, but this infection might get complicated due to the spread of organism to kidney, spleen, bone marrow, central nervous system and other organs including bone. The standard conventional methods of diagnosis are time consuming, (4-6weeks) and are not always successful, especially in cases of extra pulmonary tuberculosis. By application of the real time PCR, M. tuberculosis genome can be detected within a day with quantitation which aids in initiating the appropriate therapy immediately.

Rubella virus is a single stranded RNA virus, and is one of the causative agent of Congenital Cataract. Rubella is known to be a Teratogen with respect to the pathogenesis of the virus- induced congenital malformation. Transplacental infection of the fetus occurs during viremia. If the mother is infected within the first 20 weeks of pregnancy, the child may be born with congenital rubella syndrome (CRS), Congenital infection of the fetus with rubella is termed as CRS, wherein a triad of organs are affected namely – Heart, eyes and Central Nervous System (CNS). The disease is diagnosed by isolation technique. Recently reverse transcriptase -PCR have been developed for the rapid detection of rubella virus and is the most sensitive and specific test for detection of rubella antigen by various groups.