Molecular Diagnostics

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Gastroenteric

Hepatitis A, the most acute form of viral hepatitis prevalent throughout the globe, is an infectious disease of the liver caused by Hepatitis A virus (HAV) transmitted through the fecal-oral route. Although many cases of the disease are asymptomatic, the general symptoms of Hepatitis A include fatigue, nausea, vomiting, abdominal discomfort, clay-colored bowel movements, loss of appetite, dark urine, joint pain, jaundice and itching. HAV belongs to family Picornaviridae with non-enveloped ss (+) RNA genome (Baltimore group IV). Only one serotype and seven different genetic groups of this virus have been described. The conventional diagnostic tools for HAV detection involve the detection of HAV-specific IgM antibodies in blood and elevated levels of alanine transferase (ALT) enzyme in blood. RT-PCR based pathogen detection is an emerging tool in the diagnostics regime and highly sensitive at detection of HAV in infected human blood samples. Our Hepatitis A detection primers amplify a unique 158-bp sequence specific for Hepatitis A virus (HAV) and is absent in other closely-related Picornaviridae members and other viruses causing hepatitis. Representative gel image showing amplification of the HAV sequence. Lane 1: DiAGSure DNA ladder; Lane 2: Positive amplification of the HAV amplicon at 158-bp; Lane 3: Negative control. Amplification plots obtained from Real-time PCR of different nanograms of Hepatitis A viral genome under in vitro conditions.
Hepatitis B virus (HBV) is the causative agent of the liver disease Hepatitis B. The virus infects the liver leading to liver cirrhosis, liver failure and hepatocellular carcinoma. Both acute and chronic cases have been reported. Hepatitis B is the most common serious liver infection in the world and is more common in young adults aged 20-50 years. According to an estimate, about 2 billion people bear serological evidence of past or present HBV infection. India has over 40 million HBV infected patients and has the second highest burden of Hepatitis B infected individuals after China. The HBV is a DNA virus with a double-stranded gapped DNA genome that replicates through an RNA intermediate (Fam. Hepadnaviridae). Among the different means of Hepatitis B diagnosis, polymerase chain reaction (PCR) has been proven to be extremely useful and a sensitive diagnostic tool. Our Hepatitis B detection service detects a conserved 140-bp region in the HBV genome specific for this virus. We are able to detect as low as 7.26 attomoles of the viral DNA under in vitro conditions. Representative gel image showing amplification of the HBV gene. Lane 1: Positive amplification at 140-bp; Lane 2: Negative control; Lane 3: 221-bp Internal control amplification (IC); Lane 4: DiAGSure DNA ladder. Real-time PCR-based Hepatitis B viral DNA amplification from infected blood sample along with internal control (IC) and negative control (NTC). CT (sample) = 18.7.
Hepatitis C is a liver inflammatory disease caused by Hepatitis C Virus (HCV). The disease can occur as acute or chronic forms. The symptoms of the disease include fever, dark urine, loss of appetite, abdominal pain, jaundice and joint pain. Complications from Hepatitis C include cirrhosis and hepatocellular carcinoma. Hepatitis C is transmitted through blood contact with HCV infected individual. About 3.9 million people in the U.S. are affected by this disease. WHO estimates that 71 million people have chronic Hepatitis C. As such, accurate diagnosis of the disease proves helpful for proper treatment of the disease. The Hepatitis C virus belongs to family Flaviviridae. The genome of the virus is a single-stranded (+) RNA (Baltimore class IV). Reverse Transcriptase-Polymerase chain reaction (RT-PCR) has been proven to be extremely useful and a sensitive diagnostic tool for the detection of RNA viruses, including HCV. This HCV detection service amplifies a unique 230-bp region in the HCV genome absent in other related RNA viruses with a detection limit of 10 viral copies. Representative gel image showing amplification of the HCV sequence. Lane 1: DiAGSure DNA ladder; Lane 2: Positive amplification at HCV 230-bp region; Lane 3: Negative control. Amplification plots obtained from Real-time PCR of different nanograms of Hepatitis C viral genome under in vitro conditions.
Hepatitis E is a viral inflammatory disease of the liver caused by Hepatitis E virus (HEV). HEV is a ss (+) non-enveloped RNA icosahedral virus (Baltimore class IV).Estimation by WHO shows that 20 millions of individuals get infected by the virus worldwide every year of which around 3.3 million cases are asymptomatic. The disease is common in resource-limited countries with scarcity of drinking water and poor sanitation conditions. The disease spreads through the oral-fecal route; i.e., by the consumption of contaminated drinking water. HEV has four genotypes of which genotypes 1 and 2 are exclusive to humans. The symptoms of the disease include fever, fatigue, loss of appetite, nausea, vomiting, abdominal pain, dark urine, joint pain, jaundice and hepatomegaly. In most cases, detection of the virus is based on the detection of specific IgM antibodies to the virus in the infected blood. Reverse Transcriptase-Polymerase chain reaction (RT-PCR) has been proven to be extremely useful and a sensitive diagnostic tool to detect the presence of HEV in human blood and stool samples. Our HEV detection service amplifies a unique 198-bp sequence specific for Hepatitis E virus (HEV), and is absent in other closely-related Caliciviridae members and other viruses causing hepatitis. The limit of HEV detection is 2.45 attomoles of the virus under in vitro conditions. Representative gel image showing amplification of the HEV sequence. Lane 1: DiAGSure DNA ladder; Lane 2: Positive amplification of the HEV amplicon at 198-bp; Lane 3: Negative control.
Typhoid fever, also known as Typhoid is a bacterial disease, which is caused by Salmonella typhi. Symptoms can vary from mild to severe, and most of the time begins after one to two months of exposure. The major symptoms include high fever over several days, weakness, constipation, abdominal pain, headache, etc. Some people also develop a skin rash, known as rose colored spots. Symptoms are very common with that of other diseases; therefore proper diagnosis is very important. Typhoid fever is spread through eating or drinking food or water, contaminated with feces of infected persons. This disease is most common in India, and children are mostly affected. Diagnosis is done by blood, bone marrow or stool culture with the widal tests, which includes use of antibody-antigen. But the widal test is time-consuming and very prone to give false positive result. However, nucleic acid (PCR) based diagnosis methods are very helpful for obtaining accurate results. We target a conserved 164-bp region in the genome of Salmonella typhi with a sensitivity of 1.8 attomoles of bacterial DNA. Representative gel image showing amplification of the S. typhi 164-bp amplified product.
Cholera is a wide spread intestinal infectious disease caused by some strains of Vibrio cholera bacterium. It is usually transmitted through fecal-oral route of contaminated water or food. Symptoms can be observed within a few hours or after five days of infection which are mainly watery diarrhea, vomiting and dehydration. It is also named as “blue death” as patient’s skin may turn bluish gray due to excessive loss of fluids. If not treated properly it can lead the patient even into coma, especially for children. Microscopy is the most widespread method to diagnose this disease across the world by stool examination, but it is not so sensitive and accurate. Polymerase Chain Reaction (PCR) is considered as the best method for diagnosis of cholera. Primers have been designed to detect a conserved 414-bp region in the genome of V. cholerae pathogen. The sensitivity of the assay is 0.04 attomoles of V. cholerae genomic DNA under in vitro conditions. Representative gel image of V. cholerae 414-bp amplicon along with DiAGSure DNA ladder (band for internal control has also been shown).
Shigellosis is an intestinal disease which is caused by the bacterium Shigella. Symptoms are generally visible after one to two days exposure which include diarrhea, fever, abdominal pain etc. sometimes the diarrhea can be bloody. In the complicated situation reactive arthritis, sepsis, seizures, hemolytic uremic syndrome can also occur. It is caused by four specific types of Shigella which includes – Shigella flexneri, S. boydii, S. dysenteriae and S. sonnei. Typically these spread by exposure to infected feces. Microscopy is the most widespread method to diagnose this disease across the world by stool examination, but it is not so sensitive and accurate. Polymerase Chain Reaction (PCR) is considered as the best way for diagnosis of the disease. Here, we target a conserved 299-bp region in the Shigella genome. The absolute sensitivity of detection is 0.02 attomoles of Shigella genomic DNA. Representative gel image of 299 bp (Shigella sp.) along with DiAGSure DNA ladder (band for internal control has also been shown).
Gastric ulcer, which is also known as stomach ulcer is a very painful sores in the lining of stomach. It is one type of peptic ulcer disease, which can affect both the stomach and small intestine. It occurs when the thick layer of mucus that protects the stomach from digestive juice is reduced. Ulcer can be easily cured but it can be severe without proper diagnosis and treatment. One of the main causes of stomach ulcer is an infection by the bacterium Helicobacter pylori. They enter our body and live in the digestive tract, and after many years can cause sores, known as ulcers. For some people, it can lead to stomach cancer without a proper diagnosis and treatment. The most common symptom with ulcer is pain or burning sensation in the middle of the abdomen between chest and belly button. Other symptoms are pain in stomach, weight loss, vomiting (sometimes with blood), anemia, dark stool, etc. We aim to detect a conserved 229-bp region in the genome of Helicobacter pylori. Our limit of detection is 0.02 attomoles of H. pylori genomic DNA. Representative gel image showing amplification of the Ulcer pathogenic genes
Amoebiasis is a very common infection which occurs in the gastro-intestinal tract of human. Any of the amoeba of the Entamoeba group is responsible for the infection, out of which Entamoeba histolytica is the most common. Sometimes it can be present with no, mild or severe symptoms. About 90% of the infected people are asymptomatic but it can become more serious. Symptoms are abdominal pain, diarrhoea and bloody diarrhoea. Sometimes the situation can be more complicated. Due to loss of blood the people may suffer from anemia. The disease occurs when the amoeba comes in contact with the cells lining the intestine. Then it secretes some substances including enzymes to destroy bacterial proteins and cell membranes which lead to ulceration of the intestine. Microscopy is the most widespread method to diagnose this disease across the world by stool examination, but it is not so sensitive and accurate. Polymerase Chain Reaction (PCR) is considered as the best way for diagnosis of this disease. Our Amoebiasis detection service detects a conserved 308-bp region in the genome of Entamoeba histolytica pathogen with a sensitivity of 0.2 attomoles of E. histolytica DNA. Representative gel image showing amplification of 308-bp (E. histolytica) along with DiAGSure DNA ladder (band for internal control has also been shown).