Deoxyribonucleic Acid Polymerase Chain Reaction Research Articles

Prevention of mother-to-child transmission of HIV service interruptions amid COVID-19 pandemic.

The coronavirus disease 2019 (COVID-19) caused global disruptions in healthcare service delivery. The prevention of mother-to-child transmission (PMTCT) of human immunodeficiency viruses (HIV) services were also interrupted, threatening the attainment of Sustainable Development Goal 3. This article describes the PMTCT service interruptions experienced during the COVID-19 pandemic in Tshwane healthcare facilities. A descriptive phenomenological design was used to explore and describe the experiences of healthcare providers offering PMTCT services during COVID-19 in the Tshwane district, Gauteng province. Purposive sampling was used to recruit participants. Data were collected through in-depth interviews with 16 participants, and Colaizzi's data analysis steps were followed in analysing the findings. Participants reported interruptions in PMTCT service delivery during the pandemic. Non-adherence to scheduled visits resulted in patients defaulting or not adhering to treatment regimens, high viral loads and mother-infant pairs' loss to follow-up. Other features of service disruption included late antenatal bookings, low client flow and delays in conducting deoxyribonucleic acid-polymerase chain reaction (DNA-PCR) testing in HIV-exposed babies. In addition, staff shortages occurred because of re-assignments to COVID-19-related activities. Study participants were psychologically affected by the fear of contracting COVID-19 and worked in a frustrating and stressful environment. Improved community-based follow-up services are critical to enhance PMTCT service outcomes and prevent infant HIV infections.Contribution:The findings may influence policymakers in developing strategies to curb HIV infections among mothers and children during pandemics.

An assessment of turnaround times of infant Deoxyribonucleic acid-Polymerase Chain Reaction testing and the associated factors in Western Kenya: A mixed methods study.

The HIV/AIDS continues being a significant global public health priority in the 21st century with social and economic consequences Mother-to-child transmission (MTCT) occurs when an HIV-infected woman passes the virus to her infant and about 90% of these MTCT infections occurs in Africa where children and infants are still dying of HIV. Early definitive diagnosis using Deoxyribonucleic acid reaction of HIV infection in infants is critical to ensuring that HIV-infected infants receive appropriate and timely care and treatment to reduce HIV related morbidity and mortality. To assess the Infant Deoxyribonucleic acid-Polymerase Chain Reaction (DNA-PCR) Turnaround Time (TAT) of dry blood spots and associated factors in Vihiga, Bungoma, Kakamega and Busia counties, in Kenya. A mixed methods study using a) retrospectively collected data from Ministry of Health Laboratory registers, Early Infant Diagnosis (EID) database from 28 health facilities and b) 9 key informant interviews with laboratory in-charges were conducted. A total of 2,879 HIV exposed babies' data were abstracted from January 2012 to June 2013. The mean TAT from specimen collection and results received back at the facilities was 46.90 days, Vihiga county having the shortest mean duration at 33.7days and Kakamega county having the longest duration at 51.7days (p = 0.001). In addition, the mean transport time from specimen collection and receipt at Alupe Kenya Medical Research Institute (KEMRI) reference Laboratory was 16.50 days. Vihiga County had the shortest transport time at 13.01 days while Busia had the longest at 18.99 days (p = 0.001). Longer TAT was due to the batching of specimens at the peripheral health facilities and hubbing to the nearest referral hospitals. The TAT for DNA-PCR specimen was 46.90 days with Vihiga County having the shortest TAT due to lack of specimen batching and hubbing. Discourage specimen batching/hubbing and support point-of-care early infant diagnosis (EID) tests.

Optimization of deoxyribonucleic acid extraction and polymerase chain reaction methods for gene detection of Toxoplasma gondii in goat milk

Abstract. Fatmawati M, Suwanti LT, Mufasirin, Subekti DT, Ekawasti F, Lastuti NDR, Lamid M, Suprihati E, Effendi MH, Al-Arif A. 2023. Optimization of deoxyribonucleic acid extraction and polymerase chain reaction methods for gene detection of Toxoplasma gondii in goat milk. Biodiversitas 24: 5905-5911. Dairy goats (Capra aegagrus hircus) are intermediate hosts that transmit Toxoplasma gondii to humans through goat milk consumption. T. gondii can invade mammary glands and secrete into milk. The method of detecting gene of the T. gondii in milk is through a Polymerase Chain Reaction (PCR) test based on specific genes. The objective of this study was to compare the efficacy methods of the T. gondii molecular assay from the extraction, master mix for PCR assay, and primer that is possible to amplify the gene of T. gondii in goat milk for PCR assay. The research method for purification used flotation using concentrated sugar. This research compared the extraction kit from Kit Extraction 1 (K1), Kit Extraction 2 (K2), and Kit Extraction 3 (K3) with the extraction method listed in the brochure. Meanwhile, for the PCR assay, three types of master mix were used, including Master Mix 1 (M1), Master Mix 2 (M2), and Master Mix 3 (M3). The specific genes used included primers B1, ROP, GRA 1, GRA 7, BAG 1, and P30. Tris borate EDTA (TBE) or Phosphate Buffer Saline Solution (PBS) as buffer milk samples gave the same PCR test results. The results show that the principle of a flotation method test using PBS can be used to prepare the milk sample. The research showed that extraction kit 1 (K1) and extraction kit 2 (K2) gave positive results for the positive control. However, in terms of price, K2 is cheaper than K1. So, to optimize DNA extraction, K2 can be used. The master mix that can be used for PCR testing of milk samples is M1 because M1 gives positive results for positive controls and negative results for negative controls. Primers for optimal results used GRA 7 and B1 with denaturation temperature of 94°C, annealing at 56°C, and an extension of 73°C 33. This research concluded that the type of DNA extraction kit affects the PCR test results for detecting T. gondii in goat milk. It is necessary to optimize the type of master mix to be used in PCR testing and primers capable of amplifying the T. gondii gene.

Early infant diagnosistesting for HIV in a hard-to-reach fishing community in Uganda.

Infants born to HIV-infected mothers are at a high risk of acquiring the infection. The World Health Organization recommends early diagnosis of HIV-exposed infants (HEIs) through deoxyribonucleic acid polymerase chain reaction (DNA PCR) and rapid HIV testing. Early detection of paediatric HIV is critical for access to antiretroviral therapy (ART) and child survival. However, there is limited evidence of the factors associated with receiving early infant diagnosis (EID) tests of the HIV testing protocol among HEIs in fishing communities in Uganda. This study established the factors associated with receiving EID tests of the HIV testing protocol among HEIs in a hard-to-reach fishing community in Uganda. A cross-sectional study was conducted among HEIs in selected healthcare facilities in Buvuma islands, Buvuma district. We obtained secondary data from mother-infant pair files enrolled in the EID program using a data extraction tool. Data were analysed using STATA Version 14. A modified Poisson regression analysis was used to determine the factors associated with not receiving the 1st DNA PCR test among HEIs enrolled in care. None of the HEIs had received all the EID tests prescribed by the HIV testing protocol within the recommended time frame for the period of January 2014-December 2016. The proportion of infants that had received the 1st and 2nd DNA PCR, and rapid HIV tests was 39.5%, 6.1%, and 81.0% respectively. Being under the care of a single mother (PR = 1.11, 95% CI: 1.01-1.23, p = 0.023) and cessation of breastfeeding (PR = 0.90, 95% CI: 0.83-0.98, p = 0.025) were significantly associated with not receiving the 1st DNA PCR. Our study revealed that none of the HEIs had received all the EID tests of the HIV diagnosis testing protocol. Receiving the 1st DNA PCR was positively associated with being an infant born to a single mother, and exclusive breastfeeding. Our findings highlight the need for the creation of an enabling environment for mothers and caregivers in order to increase the uptake of early diagnosis services for HEIs. Awareness-raising on the importance of EID should be scaled up in fishing communities. Demographic characteristics such as marital and breastfeeding status should be used as an entry point to increase the proportion of HEIs who receive EID tests.

Pantoeaananatis em Oryza sativa no Brasil

ABSTRACT: The state of Santa Catarina is the second-largest producer of rice seeds in Brazil. Research on phytopathogenicbacterias in this crop is scarce and the high frequency of panicle diseases leads to the hypothesis that seeds may be infected by bacteria. This research quantified the incidence of bacteria in the seeds, verified the bacteria viability during the storage period and characterized the associated bacteria. Seeds from the 2018/19 and 2019/20 seasons were analyzed. To check the incidence, the seeds were disinfected, plated on a nutrient agar + fungicide culture medium, and incubated for seven days at 27 °C. To assess viability, every 45 days, three cultivars stored in a processing unit were subjected to the same detection methodology. To characterize, prevalent colonies were isolated on semi-selective culture medium Pantoea genus-specific agar (PGSA), where the ones that showed growth were subjected to deoxyribonucleic acid (DNA) extraction and Polymerase Chain Reaction (PCR), DNA sequencing, and sequence comparison on GenBank. The hypersensitivity reaction (HR) in tobacco was performed using a bacterial suspension of each isolate. All seed samples had an average incidence of 83%. During storage, the seeds maintained stable bacterial viability, with an average incidence of 95% at the beginning of storage and 99% at the end of it. All isolates that grew in PGSA culture medium were identified by molecular characterization with 100% identity with two specimens of Pantoeaananatis and one of them induced RH in tobacco.

Deoxyribonucleic Acid-Polymerase Chain Reaction Status of HIV Exposed Infants in a Sub Regional Prevention of Mother-to-Child Transmission of HIV Programme during the Period 2009-2020

Deoxyribonucleic Acid-Polymerase Chain Reaction Status of HIV Exposed Infants in a Sub Regional Prevention of Mother-to-Child Transmission of HIV Programme during the Period 2009-2020

Impact of HPV vaccination on HPV infection and cervical related disease burden in real-world settings (HPV-RWS): protocol of a prospective cohort

BackgroundCervical cancer is one of the most common cancers in women and could be prevented by human papilloma virus (HPV) vaccination. Cervarix, the first available HPV vaccine, has been widely administrated to Chinese women, while little was known about its effect on the prevention and control for HPV related diseases in China. The study aims to assess the impact of Cervarix on HPV infection and cervical related diseases in real world.MethodsThis is a prospective, multi-age birth cohort study to investigate the incidence and continuous status of HPV infection, and relevant cervical diseases by exposure status (with Cervarix vaccination history or without any HPV vaccination history). It is planned to recruit 12,118 eligible women at age of 9 to 45 years from vaccination clinics or hospital outpatient clinics, and then follow up them for three years. The standard questionnaire will be used to collect information such as demographic characteristics, menstruation and obstetrical histories, history of sexual behavior, personal behavior history, history of disease and pathogen infection, medication history, and family history at baseline. After three years, the changes of these behaviors will be investigated again, and other related health status information will be retrieved from the electronic health records during the follow-up period. If available physically and legally, the cervical cancer screening will be performed, including type-specific HPV deoxyribonucleic acid (DNA) polymerase chain reaction (PCR) testing and contingent thinprep cytologic test (TCT) and colposcopy. The free cervical cancer screening will be captured and uploaded timely to the Yinzhou Regional Health Information Platform (YRHIP); therefore, the long-term outcomes of participants will be monitored.DiscussionThis prospective cohort study will assess the impact of HPV vaccine on HPV infection and related cervical diseases in women aged 9–45 years, which makes up for the lack of evidence in Chinese women. The results of this study will provide support for understanding the impact of HPV vaccination in China, and make a contribution to increasing HPV vaccination and cervical cancer screening coverage in China.Trial registrationThis study has been retrospectively registered on clinicaltrials.gov (NCT05341284) on April 22, 2022.

Use of Diverse Extraction Protocols to Decide the Integrity of Deoxyribonucleic Acid (DNA) Samples Extracted from Bovine Bone Samples of Different Ages obtained from an Abattoir in Ikorodu, Lagos State, Nigeria

Techniques for the identification human and non-human biological samples are developing at very high rates with the advent of different DNA extraction methods and polymerase chain reaction (PCR) based assays. This study aimed at using different extraction protocols to determine the integrity of DNA samples extracted from bovine bone samples of different ages collected from abattoir in Ikorodu, Lagos State, Nigeria. The DNA was extracted using CTAB, PCI protocols and a DNA kit (Quick DNA MiniPrep Plus Kit). Bovine mtDNA fragment containing the gene encoding ATPase 8 was amplified via Polymerase Chain Reaction (PCR). The PCR products were analysed on 1.8% agarose gel. It was observed that the DNA samples extracted using the PCI method at 48 h incubation time had the highest purity (1.68) and concentration (336 ng/µl) compared to other extraction methods employed in the study. However, DNA kit extracted samples had mean ± SE purity (1.52 ± 0-05) and concentration (192.25 ± 31.41 ng/µl) values that were higher than CTAB protocol values but lower than PCI protocol values. All isolated DNA samples were PCR-worthy and thus yielded PCR products which were within the expected amplicon size of 126-bp. All DNA extraction protocols employed in this study are stable and efficient for use in the identification of aged non- human bones. This study also revealed that these protocols can be used to isolate PCR amplifiable DNA from old bones.

Progressive familial intrahepatic cholestasis-2 and concomitant cytomegalovirus infection in an infant: a case report

Conjugated hyperbilirubinemia (CH) in infancy can have anatomic, infectious, auto-immune and metabolic causes. We reported a case of CH presented at 2 months and 20 days, with clinical jaundice and evidence of acute on chronic liver failure. Significant history of elder sibling death at the age of 5 months due to hepatic failure and intracranial bleed was present. Investigations revealed a normal gamma glutamyl transpeptidase (GGT) levels. Toxoplasmosis, rubella, cytomegalovirus and herpes simplex (TORCH) panel showed cytomegalovirus (CMV) immunoglobulin M (IgM), immunoglobulin G (IgG) and deoxyribonucleic acid polymerase chain reaction (DNA PCR) positive with mother’s serum CMV IgG, so she was started on oral valganciclovir in addition to other treatment for cholestasis. A clinical exome panel was also sent which showed homozygous two base-pair deletion in exon 27 of the ABCB11 gene [c.3659 3660del (p.Ser1220Ter)] suggestive of benign recurrent intrahepatic cholestasis (BRIC)/progressive familial intrahepatic cholestasis-2 (PFIC-2). The child responded initially to treatment showing reduced serum conjugated bilirubin level and near normalization of INR. However, she was re-admitted with deranged coagulation profile and rising conjugated bilirubin levels after 30 days of discharge and expired due to fulminant hepatic failure with encephalopathy at 6 months of age while she was being prepared for live donor (father) liver transplantation. We presented this case because we found evidence of CMV infection in a child with PFIC-2. The relative contribution of either aetiology needs to be ascertained in view of early recurrence of CH despite standard management protocol including oral valgancyclovir for CMV infection.

Association Of ADAM33 SNP (RS528557) Gene Polymorphism With COPD In Pakistani Population

Chronic obstructive pulmonary disease (COPD) is a major health Problem worldwide. It is currently the fourth leading cause of death with the highest morbidity and mortality throughout the world. ADAM33 has been implicated in the etiology of asthma, another obstructive pulmonary disease. Research shows that its genetic polymorphism may play a pivotal role in COPD pathophysiology; however, data is still inconclusive and no research has been done on it in Pakistan. A total of 102 subjects (51 cases + 51 controls) were recruited. Blood samples were drawn for deoxyribonucleic acid (DNA) isolation from individuals. DNA extraction and Polymerase Chain Reaction (PCR) was optimized and restriction fragment length polymorphism (RFLP) was carried out by incubation at 37οC with digesting enzyme’ Fsel’ for minor allele rs528557. Data was analyzed by using SPSS version 26.0. Data for age, pack smoking/year, frequency of exacerbation and BMI was described by mean ± SD. Alleles and genotypes were described as proportions and percentages. Comparison of the said variables between two groups was performed by Chi-Square as applicable. G allele was found in all cases (100%) and in 74.5% controls at p= <0.001. On the other hand, the frequency of minor allele C was 11.8% and 29.4% in cases and controls respectively at p=0.03. Homozygous major genotype (G/G) was 88.2%, in controls versus 70.6% in cases (p=0.09). Heterozygous genotype (G/C) was 9.2% in controls and 25.5% in cases. Similarly homozygous minor genotype (C/C) was 0.8% in controls and 3.9% in cases respectively at p=0.56. Thus, we show that G allele of rs528557 may be associated with risk of COPD in Pakistani subjects.

Role of Ayurveda treatment in the management of hepatitis B (Ubhayapatha Ashrita Swatantra Kamala)- A case report.

Hepatitis B is one of the most common causes of liver disease, and due to unawareness of the safety measures, more prone to spread. As per the World Health Organization, for the South East Asia region, its prevalence is 2%. Yellowish discoloration of the eyes, body, and urine, abdominal discomfort, and vomiting is its cardinal symptoms. In Ayurveda, this set of symptoms is known as Kamala. This case report illustrates the effectiveness of the Ayurveda treatment modality in a patient with a viral load of 3705.71 IU/ml, and the values of AST (aspartate aminotransferase) and alanine transaminase (ALT) were 140 IU/ml and 173 IU/ml, respectively. A 40-year-old female patient with a complaint of yellowish discoloration of urine, eyes, and skin with fatigue and irritability was diagnosed with Ubhayapatha Ashrita Swatantra Kamala. The patient was treated with Ayurveda drugs and Virechana Karma (therapeutic purgation). After the treatment for 13 months, the disease was cured. Improvement was observed based on hepatitis B virus deoxyribonucleic acid (DNA) polymerase chain reaction (PCR) (hepatitis B DNA PCR) (decreased from 3705.71 IU/ml to <50.0 IU/ml) and the values of AST (decreased from 140 IU/ml to 19.0 IU/ml) and ALT (decreased from 173 IU/ml to 28 IU/ml). The patient was stable and asymptomatic during the follow-up period of 4 months.

Clinico-immunological Study of HIV Infected Children Attending ART CenterA Prospective Longitudinal Study

Introduction: Human Immunodeficiency Virus (HIV) infection is a growing concern in paediatric population and large number of children are registered and treated at Antiretroviral Treatment (ART) centres across the country. Children with HIV progress more rapidly, develop more bacterial infections, suffer from neurologic developmental problems and have higher mortality than adults. So the screening and counselling of HIV positive parents and their children must be done timely. This helps the physician for starting the treatment timely. Aim: To determine the clinico-immunological profile of paediatric patients registered in ART, and to compare the immunological profile and clinical staging of paediatric patients receiving ART. Materials and Methods: This prospective longitudinal study was conducted in the ART centre of Sarojini Naidu Medical College, Agra, Uttar Pradesh, India, from October 2017 to October 2018. Total 51 children, upto the age group of 18 years, suffering from HIV/Acquired immunodeficiency syndrome (AIDS) were enrolled in the study. Diagnosis of HIV was confirmed using EnzymeLinked Immunosorbent Assay (ELISA) method (using two different antigens Comb HIV test, TRI-DOT) in children more than 18 months of age. In children less than 18 months age, diagnosis was confirmed using Deoxyribonucleic Acid (DNA) Polymerase Chain Reaction (PCR) (repeated twice with cessation of breast feeding for minimum of six weeks). Statistical analysis was done by using Statistical Package for the Social Sciences (SPSS) trial version 23.0 and simple frequency and Chi-square test was used for analysis. Results: Majority of the children (20, 39.26%) were in the age group of 5-10 years, and male: female ratio was 2.4:1. Most common presenting complaint was cough (52.94%), followed by fever (47.05%), chronic diarrhea (37.25%). Most common clinical signs seen were hepatosplenomegaly (41.17%), pneumonia (33.33%) and lymphadenopathy (31.37%). Initially the mean CD4 count was 370.31±231.5 cell/mm3 , and after starting ART mean CD4 count was 524.6±260.4 cell/mm3 . Significant improvement in CD4 count was observed in age group of 5-10 years (p-value=0.009), and 10-15 years (p-value=0.001) after six months of starting the ART. In the beginning, maximum (56.7%) children belonged to World Health Organization (WHO) clinical staging III and after starting ART maximum 40% belonged to stage II. Conclusion: ART improves symptomatology and immunological status HIV infected children, so there is need to screen the children of HIV affected parents and identify the children suffering with HIV in order to initiate ART at the earliest indication in order to improve their general health, freedom from illness and better immunological status.

PCR analysis of CMV in hematology patients

Objectives: In this study we aimed to analyze cytomegalovirus (CMV) deoxyribonucleic acid (DNA) polymerase chain reaction (PCR) results in non-transplant patients. Patients and methods: Although we do not currently perform bone marrow transplant (BMT), we conducted a retrospective analysis of CMV DNA PCR monitoring (twice-weekly) of 91 hospitalized patients (with 306 peripheral blood) (47 males, 44 females; mean age 62+2.3 year; range, 25 to 85 year). For this purpose, we reviewed CMV DNA PCR records of patients in our clinic. Results: The results of CMV DNA PCR in 91 non-transplant patients are analyzed retrospectively in this study. CMV DNA PCR test results were positive in 10 patients (a total of 17 peripheral blood) while negative in 81 patients (a total of 289 peripheral blood). We did not begin antiviral therapy in patients with positive CMV DNA PCR results. Because it is incompatible with the patients’ clinical CMV DNA PCR positive CMV infection. None of the 91 patients enrolled in the study are transplant recipients, and they did not receive alemtuzumab treatment. Three of 10 CMV DNA PCR-positive patients had multiple myeloma (MM) with renal failure. One of the MM patients was diabetic. Three patients were diagnosed with acute myeloblastic leukemia and received chemotherapy; two patients were diagnosed with immune thrombocytopenic purpura; one patient was diagnosed with chronic lymphocytic leukemia, and one patient received therapy for the diagnosis of aplastic anemia. They were given treatment protocols based on their diagnosis. None of our patients has specific clinical CMV infection clinical findings. Conclusion: We conclude that close CMV DNA PCR monitoring in non-transplant hematology patients is not cost-effective. Even though it is not recommended for other hematology patients, close monitoring of CMV DNA PCR is still performed in many clinics in Turkey, and we suspect it is still performed in clinics abroad.

An Analysis of Health Science Students’ Preparedness and Perception of Interactive Virtual Laboratory Simulation

The achievement of learning goals via laboratory practical depends on both extrinsic and intrinsic factors. They could be limited by laboratory time, incurred cost, safety, self-efficacy, inadequate prior preparation by learners, and different learning styles. Hence, virtual laboratory simulation (vLAB) may be an appropriate e-learning tool to overcome these restrictions. In this study, student’s perception of the usefulness of vLAB was determined by using deoxyribonucleic acid (DNA) gel electrophoresis and polymerase chain reaction (PCR) as case examples. The perception of Year 2 and 3 health science undergraduate students’ (N = 87) was studied using a questionnaire consisting of 12 items, rated on a 5-point Likert-scale. The attainment of learning outcomes was assessed using pre-and post-tests containing multiple-choice questions (MCQs). In addition, student’s experience and learning from the vLAB were further explored using qualitative analysis. Although there was no significant difference between the mean scores of the pre-and post-tests, results showed that all participants perceived vLAB well, with a median score of 4 (Agree) for all items in the questionnaire. It provides a meaningful learning experience and an authentic environment where students feel safe to practice what they have learnt in lectures. Moreover, vLAB facilitates individualised learning and enhances self-efficacy among students. In conclusion, vLAB prepares students for physical laboratory sessions by activating the prehension dimension of Kolb’s learning cycle, therefore complementing and strengthening the attainments of health sciences laboratory learning goals and outcomes.

Parvovirus Infection Causing Aplastic Anaemia in an Immunocompetent Patient Diagnosed by Bone Marrow Aspiration Cytology

Human Parvovirus infection is associated with transient aplastic crisis as a most common complication in chronic haemolytic anaemia patients. Very few case reports are available regarding aplastic crisis in immunocompetent patients. Here the authors are reporting a case of 17-year-old immunocompetent male that presented with fever, headache for three days. Complete blood count showed pancytopenia. Bone marrow aspiration and trephine biopsy revealed hypocellularity. Giant pro-erythroblasts with multiple cytoplasmic projections (dog ear appearance) and prominent intranuclear inclusions were seen in marrow aspirated cytology. Classical cytological findings in bone marrow aspiration raised the suspicion of human parvovirus infection. Findings were confirmed by positive Deoxyribonucleic Acid Polymerase Chain Reaction (DNA PCR) for parvovirus B19 and parvovirus Immunoglobulin M (IgM) antibody using Enzyme Linked Immunosorbent Assay (ELISA) test. Patient was managed with antibiotic therapy and was trasfused three units of blood. Complete blood count became normal after two months. This emphasises the role of keeping high index of suspicious for Parvovirus B19 infection as a cause of aplastic anaemia even in individual without underlying diseases. Secondly characteristic morphological changes in the erythroid precursors in the bone marrow aspiration is highly suggestive of Parvovirus B19 infection.

GENETIC RELATIONSHIPS AMONG FOUR MUTELID SPECIES (BIVALVIA: UNIONIDA) IN EGYPT REVEALED BY RAPD-PCR TECHNIQUE

Random amplified polymorphic deoxyribonucleic acid-polymerase chain reaction (RAPD-PCR) is considered as one of the simple and quick methods that are used to resolve the genetic relationship among species of taxa that are difficult to identify on the basis of morphological characters. Bivalves show great morphological variations that couple with relatively few constant characters rendering them a systematically difficult group, and many of them have numerous subspecies and several synonyms. Due to the high degree of morphological variability within Order Unionoida, four species: Mutela dubia, M. rostrata, Chambardia rubens, and C. letourneuxi were collected from Benha region (Qaluobiya Governorate, Egypt) to resolve their degree of genetic similarity depending on RAPD-PCR technique. Out of six primers used, only five primers “UBC476, UBC477, UBC478, UBC483, and UBC486” worked successfully. The study revealed that C. rubens and C. letourneuxi are very closely related species, as they showed closely similar bands on using the primers UBC477, UBC483, and UBC486.

The association of gene rearrangement and lymphoma diagnosis: A prospective observational study.

Introduction:To investigate the gene rearrangement and mutation of lymphoma biomarkers including (Immunoglobulin H (IgH), Immunoglobulin kappa (IGK), Immunoglobulin lambda (IGL), and TCR) in the lymphoma diagnosis.Methods and analysis:Paraffin tissue samples from 240 cases diagnosed as suspected lymphoma in the department of pathology, Deyang City People's Hospital from June 2020 to June 2021 will be enrolled. Deoxyribonucleic acid extraction and Polymerase Chain Reaction (PCR) amplification will be performed in these paraffin tissue samples. Immunoglobulin and T cell receptor (TCR) rearrangement will be analyzed by hetero-double chain gel electrophoresis and BioMed-2 standardized immunoglobulin gene rearrangement detection system. In this study protocol IGH gene rearrangement, IGK gene rearrangement, both IGH and IGL gene rearrangement, both IGH and IGK gene rearrangement, both IGK and IGL gene rearrangement, both IGH, IGK and IGL gene rearrangement, TCR gene rearrangement and positive Ig/TCR rearrangement will be analyzed.Discussion:In this study, we will use B and T cell lymphoma analysis focusing on IgH, IGK, IGL, and TCR gene rearrangement, so as to provide early guidance for the diagnosis of lymphoma. Second generation sequencing technology is helpful in the differential diagnosis of lymphoma.Trial registration:Chinese Clinical trial registry: ChiCTR2000032366.

Fecal non-aureus Staphylococci are a potential cause of bovine intramammary infection

The presence of non-aureus staphylococci (NAS) in bovine rectal feces has recently been described. Similar to other mastitis causing pathogens, shedding of NAS in the environment could result in intramammary infection. The objective of this study was to investigate whether NAS strains present in feces can cause intramammary infection, likely via teat apex colonization. During a cross-sectional study in 5 dairy herds, samples were collected from the habitats quarter milk, teat apices, and rectal feces from 25%, 10%, and 25% of the lactating cows, respectively, with a cow serving as the source of one type of sample only. Samples from clinical mastitis cases were continuously collected during the 1-year study period as well. The 6 most prevalent NAS species, Staphylococcus (S.) chromogenes, S. cohnii, S. devriesei, S. equorum, S. haemolyticus, and S. hominis, were further subtyped by random amplification of polymorphic deoxyribonucleic acid polymerase chain reaction (RAPD-PCR), when the same NAS species was present in the same herd in the three habitats. For S. chromogenes, S. cohnii, S. devriesei, and S. haemolyticus, the same RAPD type was found in rectal feces, teat apices, and quarter milk, indicating that fecal NAS can infect the mammary gland. For S. hominis and S. equorum, we were unable to confirm the presence of the same RAPD types in the three habitats.

Prevalence of endometrial tuberculosis-polymerase chain reaction (TB PCR) positive (Latent TB) in a population referred for in vitro fertilisation(IVF) in a developing country

SettingStudies have suggested a possible role of antitubercular treatment (ATT) for treating infertility in women with latent or preclinical tubeculosis (TB). ObjectiveThis study investigates the effect of latent TB on IVF outcome. Study designThis was a prospective cohort study which included women referred for assisted reproductive technique between December 2013 to February 2015. Two hundred forty-five women who consented for an endometrial biopsy for TB DNA PCR (Deoxy-ribo nucleic acid-polymerase chain reaction) in the premenstrual phase were included in this study. MethodsNested TB DNA PCR for IS6110 and MPB64 was performed on endometrial samples of the women. The effect of latent TB (positive endometrial DNA PCR) was estimated in terms of statistical difference in clinical pregnancy rate. ResultsEndometrial TB DNA PCR was positive for 19 women (7.7%). The clinical pregnancy rate was similar in both groups (28.16 vs. 26.3%; p = 0.5) without ATT. Patients with both positive and negative polymerase chain reaction (PCR) were similar in age, duration and type of infertility, and the infertility diagnosis. ConclusionPositive endometrial TB DNA PCR (latent TB) alone does not require antitubercular drugs for treatment of infertility. TB DNA PCR should not be the sole criterion for diagnosis of genital TB and starting or concluding treatment.

A microbial method for improving salt swelling behavior of sulfate saline soil by an experimental study

Services of soil environment system and safety production of pavements and railways are seriously affected and threatened by salt swelling behavior of sulfate saline soil. To improve the salt swelling, a microbial method using sulfate-reducing bacteria (SRB) was presented, based on the metabolism-reduction mechanism of SRB. First, the SRB was domesticated in an increasing sodium sulfate environment up to adapt the salinity of saturated sulfate saline soil. Then the salt swelling contrast tests of sulfate saline soil with and without SRB during the cooling process were conducted. Effects of temperature and salinity on salt swelling displacement, salt swelling reduction value, and salt swelling reduction rate of sulfate saline soil with and without SRB were investigated. The test results of X-Ray Diffraction (XRD) and Deoxyribonucleic Acid-Polymerase Chain Reaction (DNA-PCR) were used to verify and explain the reduction and reduce-swelling mechanism (RRSM) of SRB on sulfate saline soil. Test results as follows: (1) SRB strain-A resistant to a high concentration sulfate was obtained through multiple domestications, and had a significant reduction and reduce-swelling effect (RRSE) on sulfate saline soil. (2) Salt swelling displacement of sulfate saline soil with and without SRB increases with increasing salinity and decreasing temperature, but the maximum salt swelling displacement of sulfate saline soil containing SRB was less than that without SRB. (3) SRB has a significant inhibitory effect on salt swelling of sulfate saline soil. The maximum reduction rate of salt swelling was 32.96% (2% salinity) and the minimum one was 11.54% (5% salinity). (4) Low temperature and high salt concentration environment inhibited SRB activity and hindered the SSRE of SRB. The reduction rate of salt swelling by SRB decreased continuously with the increase of salinity and presented an exponential function change trend. (5) Results of the DNA-PCR and XRD have effectively verified the occurrence of the RRSE of SRB. This research proves that the microbial method using SRB to improve the salt swelling is feasible and is of great significance for sulfate saline soil treatment.