Molecular Diagnostics

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Vector Borne Diseases

Dengue is a tropical mosquito-borne disease caused by dengue virus (DENV) which is transmitted by the mosquito Aedes aegypti. The symptoms of the disease include severe pain in muscles and joints, swelling of lymph nodes, headache, fever and rash. Progression of infection may lead to Dengue Hemorrhagic Fever (DHF) – a severe form of the disease involving vascular and hemostatic abnormalities. Dengue affects various countries of the Asia, the Pacific, the Americas, Africa, and the Caribbean. In India, more than 20,000 cases of dengue are reported each year. As such, accurate diagnosis of the disease proves helpful for proper treatment of the disease. The dengue virus belongs to family Flaviviridae which also includes West Nile virus, Japanese Encephalitis virus, Yellow Fever virus and Zika virus. 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 DENV. Our Dengue Detection service involves semi-quantitative RT-PCR based detection of a conserved DENV 511-bp sequence in the D1/D2 region using gene-specific primers that can detect as low as 10 copies of viral RNA under in vitro conditions. PCR-based detection is emerging as a highly sensitive diagnostic tool for the detection of pathogen in a wide array of clinical samples.

Representative gel image showing amplification of the DENV D1/D2 sequence. Lane 1: Positive amplification at 511-bp; Lane 2: Negative control; Lane 3: DiAGSure DNA ladder.
Chikungunya is a tropical mosquito-borne disease caused by Chikungunya virus (CHIKV). The symptoms of the disease include fever and acute joint pain along with nausea, rash, headache, and fatigue. The symptoms are often similar to dengue and zika. Over 60,000 cases of Chikungunya were reported in India in 2017. As such, accurate diagnosis of the disease proves helpful for proper treatment of the disease. The chikungunya virus belongs to family Togaviridae. 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 CHKV. We target RT-PCR based detection of a conserved CHIKV 212-bp sequence using gene-specific primers.

Representative gel image showing amplification of the CHIKV sequence. Lane 1: Positive amplification at CHIKV 212-bp region; Lane 2: Negative control; Lane 3: DiAGSure DNA ladder.

RT-qPCR from different nanograms (as indicated) of Chikungunya RNA
Malaria is a mosquito-borne infectious disease caused by Plasmodium parasite. It is mainly caused by infection with four Plasmodium species: Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale. Out of which P. falciparum and P. vivax is the most common and P. falciparum is most deadly. Although P. falciparum traditionally accounts for the majority of deaths, recent evidence suggests that P. vivax malaria is associated with potentially life-threatening conditions about as often as with a diagnosis of P. falciparum infection. If not treated within 24 hours, P. falciparum malaria can progress to severe illness often leading to death. Early diagnosis and treatment of malaria reduces disease and prevents deaths. It also contributes to reducing malaria transmission. We target amplification of a conserved 235-bp region in the genome of Plasmodium pathogen. In case of positive amplification, we further move towards speciation of P. vivax and P. falciparum.

Our Kala azar in-vitro detection service detects the presence of the flagellate protozoan Leishmania donovani in human blood samples. Kala Azar is a slow progressing parasitic disease and one of the most neglected tropical diseases. Leishmania donovani infects mostly reticulo-endothelial system, liver, bone marrow etc. Without proper diagnosis and treatment, Kala Azar is associated with high mortality. GDL’s Kala azar detection service is based on semi-quantitative end-point PCR based detection of a conserved Leishmania specific 116 bp region in the Leishmania genome using gene-specific primers. Representative gel image showing amplification of the Leishmania gene. Lane 1: Positive amplification at 116-bp; Lane 2: Negative control (NTC); Lane 3: 221-bp Internal control amplification; Lane 4: DiAGSure DNA ladder.
Japanese Encephalitis Virus (JEV) is an arbovirus of the family Flaviviridae. It propagates in a zoonotic cycle involving pigs, ardeid birds and Culex mosquitoes. Humans are accidental/dead end hosts of JEV infection because they cannot sustain high viral titers, though 1 in 250 JEV infections develop into encephalitis. As the latent period or asymptomatic phase of the infection ranges in days, specific and fast diagnosis of JEV infection is necessary. Our molecular JEV detection tool involves semi-quantitative RT-PCR based detection of a conserved JEV 488-bp sequence in the JEV genome using gene-specific primers. Representative gel image of positive amplification from clinical samples. Lane 1:.DiAGSure DNA ladder; Lane 2: JEV amplification (488bp); Lane 3: Negative control.
Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii. In adults, infections with toxoplasmosis usually cause no obvious symptoms. Sometimes, people may have a few weeks or months of mild, flu like illness, such as muscle aches and tender lymph nodes. Eye problems may develop in a small number of people.This disease is usually spread by eating poorly cooked food, exposure to infected cat feces, and from a mother to a child during pregnancy if the mother becomes infected. Rarely, the disease is spread by blood transfusion. It is not otherwise spread between people. The parasite is only known to reproduce sexually in the cat family. However, it can infect most types of warm-blooded animals, including humans. Up to half of the world's population is infected by toxoplasmosis, but have no symptoms. Our Toxoplasmosis detection system detects a conserved 132-bp region in the genome of T.gondii pathogen with a sensitivity of 1.8 attomoles of the pathogen genomic DNA. Representative gel image showing amplification of the T. gondii pathogen gene.
Nipah is a member of family Paramyxoviridae with a ss-(-)-RNA genome placing it under Baltimore group V. Fruit bats (Fam. Pteropodidae) serve as natural hosts for Nipah virus. Initially identified in 1999, this zoonotic virus can spread through contaminated food and directly between people. The initial symptoms of the disease include fever, headaches, myalgia, vomiting and sore throat which, if left untreated, eventually lead to acute respiratory infection, and fatal encephalitis. No treatment or vaccine for the infection is currently available and the primary treatment involves supportive care. Although the first outbreak of Nipah was reported in Malayasia, several cases of Nipah were reported in India and Bangladesh. We aim to detect a unique 127-bp sequence specific for Nipah Virus and is absent in other closely-related Paramyxoviridae members. Representative gel image showing amplification of the Nipah virus sequence. Lane 1: DiAGSure DNA ladder; Lane 2: Positive amplification of the Nipah virus amplicon at 127-bp; Lane 3: Negative control.
Zika is a viral disease spread by Aedes mosquito. It was first identified in Uganda to occur in monkeys wherefrom it spread to humans crossing the species barrier. The disease is now reported to occur globally. Although most cases of zika infection are asymptomatic, the common symptoms generally include mild and include fever, rash, conjunctivitis, muscle and joint pain, malaise or headache. The symptoms typically last for 2-7 days. Zika virus can be transmitted to the fetus from an infected mother that lead to microencephaly and other congenital disorders in the new-born, often referred to as congenital Zika syndrome. The icosahedral virus belongs to Flaviviridae (Baltimore Group IV) and its genome is a ss-(+)-RNA of about 10kb in length encoding structural and non-structural (NS) proteins. Zika virus infection can be diagnosed by laboratory tests of blood and other body fluids like urine or serum. RT-PCR based detection of the virus efficiently detects the presence of viral RNA in infected human blood and other body fluids. Our primers amplifies a unique 350-bp sequence specific for Zika virus and is absent in other closely-related Flaviviridae members. The presence of the 350-bp band from the PCR product using Zika-specific primers will indicate the presence of Zika virus in the clinical sample. Our Zika virus Detection service can detect up to 3 attomoles of the viral RNA under in vitro conditions. Representative gel image showing amplification of the Zika sequence. Lane 1: DiAGSure DNA ladder; Lane 2: Positive amplification of the Zika amplicon at 350-bp; Lane 3: Negative control.
It is a virus that causes fever, symptoms similar to those of flu, and acute encephalitis (inflammation of the brain). Chandipura virus was first isolated in 1965 in a village in Maharashtra State, India. Since then, the virus has been reported in adjoining states in central India. The likely vector (carrier) of the virus is the female phlebotomine sandfly. The virus has been detected in sandflies in Senegal and Nigeria as well as in India. In 2003 Chandipura virus was responsible for an outbreak in southern India in which 329 children developed acute encephalitis and 183 died. The disease progressed rapidly from an influenza-like illness to coma and death. Chandipura virus is a member of the Vesiculovirus genus of the family Rhabdoviridae. This virus should be considered as an important emerging pathogen. Representative gel image of positive amplification from clinical samples. Lane 1:.DiAGSure DNA ladder; Lane 2: Chandipura virus amplification (186-bp); Lane 3: Negative control.