Ecoli Dx: PCR Kits for Clinical Diagnostics and Human Genome Testing
Ecoli Dx is a Czech biotechnology company. They provide about 200 different types of AmpliSens® PCR kits for clinical diagnostics and human genome testing. The kits are designed according to laboratory standards for PCR and Real-Time PCR detection.
The PCR diagnostic kits have a high sensitivity and specificity and a very reasonable price. Most of the kits are for in vitro diagnostics and have a CE-IVD certificate.
Next to Real-Time or Fluorescent End-Point Detection Ecoli Dx offers MultiPlex Real-Time/FEP PCR kits which allow the simultaneous detection of multiple targets in a single reaction well.
This increases the speed of detection. Further advantages of using Multiplex PCR are more information with less sample, higher throughput and cost savings (e.g., fewer dNTPs, enzymes, and other consumables).
Ecoli Dx offers a full range of IVD and RUO assays focusing on the detection of infectious pathogens and diseases:
Sexually Transmitted Infections
Sexually transmitted disease (STD), also known as a sexually transmitted infection (STI) or venereal disease (VD), is an illness that has a significant risk of transmission between humans by means of human sexual behavior. While in the past these illnesses have mostly been referred to as STDs or VD, in recent years the term sexually transmitted infections (STIs) has been preferred, as it has a broader range of meaning; a person may be infected, and may potentially infect others, without having a disease.
Some STIs can also be transmitted via the use of IV drug needles after its use by an infected person, as well as through childbirth or breastfeeding. STI is a broader term than STD. Infection is a colonization by a parasitic species, which may not cause any adverse effects. In a disease the infection leads to impaired or abnormal function. In either case, the condition may not exhibit signs or symptoms. Increased understanding of infections like HPV, which infects most sexually active individuals, but cause disease in only a few has led to increased use of the term STI.
Many infectious diseases, including the common cold, influenza, pneumonia and most others that are transmitted person-to-person can also be transmitted during sexual contact if one person is infected. However, even though these diseases may be transmitted during sex, they are not considered STDs. MultiPlex Real-Time PCR technology allows using primers and probes for several (for up to 5) DNA targets in one tube. Amplification products identification runs for each DNA target on a different optical channel.
The sensitivity of these tests is not affected by changing the number of infections. Each mono- and multiplex PCR kit contains independent Internal Control (IC) for determination of DNA extraction efficiency and PCR process. Presence of the Internal Control signal/band shows, that the DNA extraction process and amplification steps were sufficient for significant results interpretation.
Human Papillomavirus Infections
Human papillomaviruses (HPVs) are a group of more than 150 related viruses. They are called papillomaviruses because certain types may cause warts or papillomas, which are benign (noncancerous) tumors. Some HPVs, such as those that cause common warts that grow on hands and feet, do not spread easily. However, more than 40 HPV types are sexually transmitted and these HPVs spread very easily through genital contact.
Some types of sexually transmitted HPVs cause cervical cancer and other types of cancer. These are called high-risk, oncogenic, or carcinogenic HCR HPVs (about 13 types). Other sexually transmitted types of HPV do not appear to cause cancer and are called low-risk HPVs (LCR HPVs).
Although genital HPV infections are very common, most occur without any symptoms and go away without any treatment within a few years. However, some HPV infections can persist for many years. Persistent infections with high-risk HPV types can cause cell abnormalities. If untreated, areas of abnormal cells (lesions) can in some cases develop into cancer.
Some types of sexually transmitted low-risk HPVs cause warts to appear on or around the genitals or anus. Most genital warts are caused by two HPV types, HPV-6 and HPV-11. Warts may appear within several weeks after sexual contact with a person who is infected with HPV, or they may take months or years to appear, or they may never appear.
Hepatitis Viruses Infections
Hepatitis A Virus
Hepatitis A is an acute infectious disease of the liver caused by the hepatitis A virus (HAV), an RNA virus, usually spread the fecal-oral route, transmitted person to person, by ingestion of contaminated food or water or through direct contact with an infectious person. HAV only causes acute hepatitis and is not associated with chronic liver disease. Most individuals infected with HAV develop non-specific constitutional signs and symptoms followed by gastrointestinal symptoms. Symptoms include fever, malaise, anorexia, nausea, abdominal discomfort, dark urine and jaundice. The disease course typically lasts less than 2 months. In rare cases, HAV can cause severe cases of fulminant hepatitis with fatal outcomes in otherwise healthy adults.
Hepatitis B Virus
Hepatitis B virus (HBV) is divided into four major serotypes (adr, adw, ayr, ayw) based on antigenic epitopes present on its envelope proteins and into eight genotypes (labeled A through H) according to overall nucleotide sequence variation of the genome. The genotypes have a distinct geographical distribution and are used in tracing the evolution and transmission of the virus. Differences between genotypes affect the disease severity, course and likelihood of complications and response to treatment and possibly vaccination. A possible new „I“ genotype has been described, but acceptance of this notation is not universal. Different genotypes may respond to treatment in different ways.
HBV is one of a few known non-retroviral viruses which use reverse transcription as a part of its replication process. Hepatitis B is an infectious illness that infects the liver and causes an inflammation called hepatitis. Transmission of HBV results from exposure to infectious blood or body fluids such as semen and vaginal fluids, while viral DNA has been detected in the saliva, tears and urine of chronic carriers with high titer DNA in serum. Perinatal infection is a major route of infection in endemic countries. Other risk factors for developing HBV infection includes working in a health care setting, transfusions and dialysis. Acute infection with HBV is associated with acute viral hepatitis - an illness that begins with general ill-health, loss of appetite, nausea, vomiting, body aches, mild fever, dark urine and then progresses to development of jaundice. It has been noted that itchy skin has been an indication of a possible symptom of all hepatitis virus types. The illness lasts for a few weeks and then gradually improves in most affected people. A few patients may have more severe liver disease (fulminant hepatic failure) and may die as a result. The infection may be entirely asymptomatic and may go unrecognized. Chronic infection with HBV may be either asymptomatic or may be associated with a chronic inflammation of the liver, leading to cirrhosis over a period of several years. This type of infection dramatically increases the incidence of hepatocellular carcinoma.
Hepatitis C Virus
Hepatitis C virus is a small, enveloped, single-stranded, positive-sense RNA virus. It is the only known member of the hepacivirus genus in the family Flaviviridae. There are six major genotypes of the hepatitis C virus, which are indicated numerically - genotype 1 etc.). Based on the NS5 gene there are three major and eleven minor genotypes. HCV genotype matters because it can affect how successful a person's hepatitis C treatment will likely be and how long the hepatitis C medication will need to be taken.
Hepatitis C is an infectious disease primarily affecting the liver, caused by the HCV. HCV is transmitted by blood-to-blood contact. In developed countries, it is estimated that 90% of persons with chronic HCV infection were infected through transfusion of unscreened blood or blood products or via injecting drug use or sexual exposure. The infection is often asymptomatic, but chronic infection can lead to scarring of the liver and ultimately to cirrhosis, which is generally apparent after many years. In some cases, those with cirrhosis will go on to develop liver failure or other complications, including liver cancer or life-threatening esophageal varices and gastric varices. During the first 12 weeks after infection with HCV, most people suffer no symptoms. For those who do, the main manifestations of acute infection are generally mild and vague and rarely point to a specific diagnosis of hepatitis C. Symptoms of acute HCV infection include decreased appetite, fatigue, abdominal pain, jaundice, itching and flulike symptoms.
HCV is usually detectable in the blood by PCR within one to three weeks after infection and antibodies to the virus are generally detectable within 3 to 15 weeks. Liver enzyme tests show variable ALT/ALS elevation. Periodically, they might show normal results. Usually, prothrombin and albumin results are normal, but may become abnormal, once cirrhosis has developed. The levels of elevation of liver tests do not correlate well with the amount of liver injury on biopsy. Viral genotype and viral load also do not correlate with the amount of liver injury. Liver biopsy is the best test to determine the amount of scarring and inflammation. The natural course of chronic hepatitis C varies considerably from one person to another. Although almost all people infected with HCV have evidence of inflammation on liver biopsy, the rate of progression of liver scarring (fibrosis) shows significant variability among individuals. Once chronic hepatitis C has progressed to cirrhosis, signs and symptoms may appear that are generally caused by either decreased liver function or increased pressure in the liver circulation, a condition known as portal hypertension. Possible signs and symptoms of liver cirrhosis include accumulation of fluid in the abdomen, bruising and bleeding tendency, varices (especially in the stomach and esophagus), jaundice and syndrome of cognitive impairment known as hepatic encephalo pathy (HE). HE is due to the accumulation of ammonia and other substances normally cleared by a healthy liver.
Hepatitis D Virus
Hepatitis D is caused by a small circular enveloped RNA virus. HDV is considered to be a subviral satellite because it can propagate only in the presence of the HBV. Transmission of HDV can occur via simultaneous infection with HBV (coinfection) or superimposed on chronic hepatitis B or hepatitis B carrier state (superinfection). Both superinfection and coinfection with HDV results in more severe complications than with HBV alone.
Hepatitis G Virus
Hepatitis G is a form of liver inflammation caused by HGV from the Flaviviridae family. It is known that transfused blood containing HGV has caused some cases of hepatitis. For this reason, patients with hemophilia and other bleeding conditions who require large amounts of blood products are at risk of hepatitis G. Also, at risk are patients with kidney disease with blood exchange by hemodialysis.
HIV and HIV-Associated Infections
HIV stands for ‚human immunodeficiency virus‘. HIV is a virus (of the type called retrovirus) that infects cells of the human immune system (mainly CD4 positive T cells and macrophages), and destroys or impairs their function. Infection with this virus results in the progressive deterioration of the immune system. Within the retrovirus family, HIV belongs to a subgroup known as lentiviruses, or „slow“ viruses. Lentiviruses are known for having a long time period between initial infection and the beginning of serious symptoms. Similar versions of HIV infect other non-human species, such as feline immunodeficiency virus (FIV) in cats and simian immunodeficiency virus (SIV) in monkeys and other non-human primates. The immune system is considered deficient when it can no longer fulfill its role of fighting off infections and diseases.
Immunodeficient people are more susceptible to a wide range of infections, most of which are rare among people without immune deficiency. Infections associated with severe immunodeficiency are known as ‚opportunistic infections‘, because they take advantage of a weakened immune system. Some people at the time of seroconversion develop “Acute retroviral syndrome” which is a glandular fever-like illness with fever, rash, joint pains and enlarged lymph nodes. Seroconversion refers to the development of antibodies to HIV and usually takes place between 1 and 6 weeks after HIV infection has happened.
Whether HIV infection causes initial symptoms or not, an HIV infected person is highly infectious during this initial period and can transmit the virus to another person. The only way to determine whether HIV is present in a person's body is by testing for HIV antibodies, DNA or RNA. After HIV has caused progressive deterioration of the immune system, increased susceptibility to infections may lead to symptoms. Primary HIV infection - may be asymptomatic or experienced as Acute retroviral syndrome. Clinical stage 1 - asymptomatic or generalized swelling of the lymph nodes Clinical stage 2 - minor weight loss, mucocutaneous manifestations and recurrent upper respiratory tract infections Clinical stage 3 - includes unexplained chronic diarrhea, unexplained persistent fever, oral candidiasis or leukoplakia, severe bacterial infections, pulmonary tuberculosis, and acute necrotizing inflammation in the mouth. Some people with clinical stage 3 have AIDS. Clinical stage 4 - includes 22 opportunistic infections or cancers related to HIV. All people with clinical stage 4 have AIDS.
Respiratory Infections
Respiratory tract infection refers to any of a number of infectious diseases involving the respiratory tract. An infection of this type is normally further classified as an upper respiratory tract infection (URI) or a lower respiratory tract infection (LRI). Lower respiratory infections, such as pneumonia, tend to be far more serious conditions than upper respiratory infections, such as the common cold. URIs represents the most common acute illness evaluated in the outpatient setting and is a nonspecific term used to describe acute infections involving the nose, paranasal sinuses, pharynx, larynx, trachea and bronchi. URIs range from the common cold - typically a mild, self-limited, catarrhal syndrome of the nasopharynx - to life-threatening illnesses such as epiglottitis. Symptoms of URIs can include cough, sore throat, runny nose, nasal congestion, headache, low-grade fever, facial pressure and sneezing. Influenza is a systemic illness that involves the upper respiratory tract and should be differentiated from other URIs. LRIs are generally more serious than URIs. LRIs are the leading cause of death among all infectious diseases. The two most common LRIs are bronchitis and pneumonia. Influenza affects both the upper and lower respiratory tracts, but more dangerous strains such as the highly pernicious H5N1 tends to bind to receptors deep in the lungs. Viruses cause most URIs, with rhinovirus, parainfluenza virus, coronavirus, adenovirus, respiratory syncytial virus, coxsackievirus and influenza virus. Human metapneumovirus is a newly discovered agent causing URIs. Group A beta-hemolytic streptococci (GABHS) cause 5% to 10% of cases of pharyngitis in adults. Other less common causes of bacterial pharyngitis include group C beta-hemolytic streptococci, Corynebacterium diphtheriae, Neisseria gonorrhoeae, Arcanobacterium haemolyticum, Chlamydia pneumoniae, Mycoplasma pneumoniae and Herpes simplex virus. Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis are the most common organisms that cause the bacterial superinfection of viral acute sinusitis. Less than 10% of cases of acute tracheobronchitis are caused by Bordetella pertussis, B. parapertussis, M. pneumoniae or C. pneumoniae.
Herpes-Virus Infections
The Herpesviridae are a large family of DNA viruses, that cause diseases in humans. The family name is derived from the Greek word herpein („to creep“), referring to the latent, recurring infections typical of this group of viruses. Herpesviruses all share a common structure - all herpes viruses are composed of relatively large ds linear DNA encoding 100-200 genes and all herpes viruses are nuclear-replicating - the viral DNA is transcibed to RNA within the infected cell's nucleus. Infection is initiated when a viral particle contacts a cell. Following binding, the virion is internalized and dismantled, allowing viral DNA to migrate to the cell nucleus, where replication of viral DNA and transcription of viral genes occurs. During symptomatic infection, infected cells transcribe lytic viral genes. In some host cells, a small number of viral genes termed latency-associated transcript (LAT) accumulate instead. In this fashion, the virus can persist in the cell (and thus the host) indefinitely. While the primary infection is often accompanied by a self-limited period of clinical illness, long-term latency is symptom-free. Reactivation of latent viruses has been implicated in a number of diseases. Following activation, transcription from latency-associated LAT to multiple lytic genes lead to enhanced replication and virus production. Clinically, lytic activation is often accompanied by the emergence of non-specific symptoms such as low-grade fever, headache, sore throat, malaise and rash as well as clinical signs such as swollen or tender lymph nodes and immunological findings such as reduced levels of natural killer cells. In this family, there are eight human herpes-viruses: Herpes Simplex virus type 1 and 2, VZV, CMV, EBV and HHV-6, HHV-7, HHV-8.
TORCH Infections
TORCH complex (also known as STORCH, TORCHES or the TORCH infections) is a medical acronym for a set of perinatal infections (i.e. infections that are passed from a pregnant woman to her fetus). The TORCH infections can lead to severe fetal anomalies or even fetal loss. They are a group of viral, bacterial, and protozoan infections that gain access to the fetal bloodstream transplacentally via the chronic villi. Hematogenous transmission may occur at any time during gestation or occasionally at the time of delivery via maternal-to-fetal transfusion.
The TORCH complex was originally considered to consist of four conditions, with the „TO“ referring to „Toxoplasma“. The four-term form is still used in many modern references, and the capitalization „TORCH“ is sometimes used in these contexts. Alternatively, the „O“ is redefined as „other“, and the acronym is spelled out as follows:
- T – Toxoplasmosis/Toxoplasma gondii
- O – Other infections (see below)
- R – Rubella
- C – Cytomegalovirus
- H – Herpes simplex virus
The „other agents“ included under O are Hepatitis B, Coxackievirus, Syphilis, Varicella-Zoster virus, HIV and Parvovirus B19.
Purulent Septic Infections
Purulent infections are characterized by purulent inflammation of tissues that arise in the implementation of pyogenic bacteria, most commonly Streptococcus, Staphylococcus, more rarely Pseudomonas or E. coli. For some common infections local centers of suppuration (glanders, bubonic plague, cutaneous anthrax) are typical. Purulent infection can develop in the form of the disease (furuncle, carbuncle, erysipelas, osteomyelitis, etc.), or as a complication of the wound. In some cases, purulent focus can disappear spontaneously or may be disposed of after a simple intervention, in others requires a complex operation. Generalization of the purulent process may lead to the development of general purulent infection, i.e., sepsis. Purulent infections are very often resistant to antibiotics.
Neuro Infections
Enterovirus
Enterovirus enters the body through the gastrointestinal tract and thrives there, often moving on to attack the nervous system. Enteroviruses can be found in the respiratory secretions or stool of an infected person. Most people infected with Enterovirus have no disease at all. Infected people who become ill usually develop either mild upper respiratory symptoms, a flu-like illness with fever and muscle aches, or illness with rash. Less commonly, some people have aseptic or viral meningitis. Rarely, a person may develop an illness that affects the heart or the brain or causes paralysis. Enterovirus infections are suspected to play a role in the development of juvenile-onset diabetes mellitus.
Poliovirus
Poliovirus is a human enterovirus and member of the family of Picornaviridae. The genome is a single-stranded RNA genome and because of its short genome and its simple composition is poliovirus widely regarded as the simplest significant virus. There are 3 serotypes of Poliovirus, PV1, PV2 and PV3. PV1 is the most common form encountered in nature, however all three forms are extremely infectious. Poliovirus infection occurs via the fecal-oral route. The virus is shed in the feces of infected individuals.
In 95% of cases only a primary, transient presence of viremia occurs and the poliovirus infection is asymptomatic. In about 5% of cases, the virus spreads and replicates in other sites such as brown fat, reticuloendothelial tissue and muscle. The sustained viral replication causes secondary viremia and leads to the development of minor symptoms such as fever, headache and sore throat. The paralytic poliomyelitis occurs in less than 1 % of poliovirus infections. The paralytic disease occurs when the virus enters the central nervous system and replicates in motor neurons within the spinal cord, brain stem, or motor cortex, resulting in the selective destruction of motor neurons leading to temporary or permanent paralysis. In rare cases, paralytic poliomyelitis leads to respiratory arrest and death. In cases of the paralytic disease, muscle pain and spasms are frequently observed prior to weakness and paralysis.
Listeria monocytogenes
L. monocytogenes is one of the most virulent food-borne pathogens. It is the third-most-common cause of meningitis in newborns. When the infection is not invasive, any illness as a consequence of infection is termed febrile gastroenteritis. The manifestations of listeriosis include septicaemia, meningitis, encephalitis, corneal ulcer, pneumonia, and intrauterine infections in pregnant women, which may result in abortion or stillbirth. Surviving neonates of fetomaternal listeriosis may suffer granulomatosis infantiseptica and may suffer from physical retardation. Influenza-like symptoms, including persistent fever, usually precede the onset of the disorders.
Intestinal Infections
Helicobacter pylori
Helicobacter pylori is a bacterium that causes chronic inflammation of the inner lining of the stomach (gastritis) in humans. It causes a chronic low-level inflammation of the stomach lining and is strongly linked to the development of duodenal and gastric ulcers, stomach cancer. H. pylori infection is most likely acquired by ingesting contaminated food and water and through person to person contact. Over 80 percent of individuals infected with the bacterium are asymptomatic.
Norovirus
Norovirus, sometimes referred to as the winter vomiting bug, is the most common cause of gastroenteritis. Infection is characterized by non-bloody diarrhea, vomiting, and stomach pain. Fever or headaches may also occur. Symptoms usually develop 12 to 48 hours after being exposed, and recovery typically occurs within 1 to 3 days. Complications are uncommon, but may include dehydration, especially in the young, the old, and those with other health problems.
The virus is usually spread by the fecal–oral route. This may be through contaminated food or water or person-to-person contact. It may also spread via contaminated surfaces or through air from the vomit of an infected person. Risk factors include unsanitary food preparation and sharing close quarters.
Especially Dangerous and Feral Herd Infections
- Vibrio cholerae
- Bacillus anthracis
- Brucella species
- Dengue fever virus
- Leptospira species
- Borrelia burgdorferi sensu lato
- Tick-borne encephalitis virus
- West Nile fever virus
- Crimean-Congo hemorrhagic fever virus
- Yersinia pestis
- Coxiella burnetii
- Ebola Zaire virus
- Zika virus
- Yellow fever virus
- Rickettsia
Human Genetics
Leucosis Quantum M-bcr
Chronic myelogenous (or myeloid) leukemia (CML), also known as chronic granulocytic leukemia (CGL), is a cancer of the white blood cells. It is a form of leukemia characterized by the increased and unregulated growth of predominantly myeloid cells in the bone marrow and the accumulation of these cells in the blood. CML is a clonal bone marrow stem cell disorder in which proliferation of mature granulocytes (neutrophils, eosinophils, and basophils) and their precursors are the main finding. It is a type of myeloproliferative disease associated with a characteristic chromosomal translocation called the Philadelphia chromosome. CML is now largely treated with tyrosine kinase inhibitors (TKIs), such as imatinib, dasatinib or nilotinib, which has led to dramatically improved survival rates since their introduction in the last decade.
CML was the first malignancy to be linked to a clear genetic abnormality, the chromosomal translocation known as the Philadelphia chromosome. In this translocation, parts of two chromosomes (the 9th and 22nd by conventional karyotypic numbering) switch places. As a result, part of the BCR („breakpoint cluster region“) gene from chromosome 22 is fused with the ABL gene on chromosome 9. This abnormal „fusion“ gene generates a protein of p210 or sometimes p185 weight (p210 is short for 210 kDa protein, a shorthand used for characterizing proteins based solely on size). Because abl carries a domain that can add phosphate groups to tyrosine residues (a tyrosine kinase), the bcr-abl fusion gene product is also a tyrosine kinase.
The fused BCR-ABL protein interacts with the interleukin 3 beta (c) receptor subunit. The bcr-abl transcript is continuously active and does not require activation by other cellular messaging proteins. In turn, BCR-ABL activates a cascade of proteins that control the cell cycle, speeding up cell division. Moreover, the BCR-ABL protein inhibits DNA repair, causing genomic instability and making the cell more susceptible to developing further genetic abnormalities. The action of the BCR-ABL protein is the pathophysiologic cause of chronic myelogenous leukemia.
With improved understanding of the nature of the BCR-ABL protein and its action as a tyrosine kinase, targeted therapies (the first of them was imatinib mesylate) that specifically inhibit the activity of the BCR-ABL protein, have been developed. These tyrosine kinase inhibitors can induce complete remissions in CML, confirming the central importance of bcr-abl as the cause of CML.
Clinically, leukemia is manifested in three distinct phases: chronic, accelerated, and blast. Most patients present in the chronic phase, a stage that is typically indolent in nature. Mature granulocytes are found, but patients typically have an increase in the number of myeloid progenitor cells found in the blood. Left untreated, the disease progresses to an accelerated phase followed by blast crisis, which is inevitably fatal. During blast phase, hematopoietic differentiation is blocked and blast cells accumulate in the bone marrow and peripheral blood. Expression of BCR-ABL onco-proteins in hematopoietic cells induces resistance to apoptosis, growth factor independence and leukemogenesis.
Additional Kits
- Nucleic Acid Extraction Kits
- Reverse Transcription
- Transport and Storage Media
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