Maternal malaria parasitemia was assessed every 4 weeks throughout pregnancy, beginning at enrollment. The presence of malaria parasites in maternal peripheral blood was evaluated using LAMP kits (Eiken Chemical), as previously described (27 (link), 28 (link)). LAMP assays were batch analyzed at the end of the trial and were not used to inform treatment (asymptomatic parasitemia is not treated per current Ugandan standard of care). Placental tissue was processed for histopathologic evidence of PM, as previously described (27 (link), 29 ), and placental blood and cord blood were tested for the presence of malaria parasites by both LAMP and microscopy.
Lamp kit
Manufactured by Eiken Chemical
Sourced in Japan
The LAMP kit is a laboratory equipment designed for Loop-Mediated Isothermal Amplification (LAMP) technology. The LAMP kit provides the necessary reagents and components for performing isothermal nucleic acid amplification. The core function of the LAMP kit is to facilitate the rapid and sensitive detection of target genetic sequences.
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Lab products found in correlation
5 protocols using lamp kit
1
Maternal Malaria Assessment in Pregnancy
2
Malaria Surveillance in Pregnant Women
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Mothers were evaluated throughout pregnancy for Plasmodium parasitaemia beginning at enrolment (12–20 weeks of gestational age), and additionally with routine monthly surveillance testing peripheral blood via loop-mediated isothermal amplification (LAMP) kits (Eiken Chemical) which detect Plasmodium DNA [18 (link), 19 (link)]. During febrile episodes mothers were evaluated with blood microscopy, and if positive, treated per local guidelines for clinical malaria, as previously described [18 (link)].
At the time of delivery, maternal peripheral blood, placental blood and cord blood was tested for parasitaemia by both LAMP and microscopy. Placental tissue was processed for histopathologic evidence of malaria infection, determined by standardized placental malaria histopathology criteria as previously described [18 (link), 20 (link), 21 (link)].
At the time of delivery, maternal peripheral blood, placental blood and cord blood was tested for parasitaemia by both LAMP and microscopy. Placental tissue was processed for histopathologic evidence of malaria infection, determined by standardized placental malaria histopathology criteria as previously described [18 (link), 20 (link), 21 (link)].
3
Malaria Diagnosis via Giemsa Staining
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Blood smears were stained with 2% Giemsa and read by experienced laboratory technologists. A blood smear was considered negative when the examination of 100 high-power fields did not reveal asexual parasites. For quality control, all slides were read by a second microscopist, and a third reviewer would settle any discrepant readings. DBS were tested for the presence of malaria parasites using a loop-mediated isothermal amplification (LAMP) kit (Eiken Chemical, Japan).
4
LAMP and Turbidity for FAdV Detection
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The DNAs or RNAs of FAdVs (1–7) and the control strains (9–16) were extracted according to the instructions of the DNA/RNA extraction kit (BioTek Co., China). RNA specimens were reverse-transcribed to cDNA using the RT Mix Kit (Takara, China).
The LAMP kit was obtained from Eiken Chemical Co., Japan. The LAMP reaction system (25 μL) contained 12.5 μL of 2 × buffer, 1 μL of enzyme mixture, FIP and BIP (40 pmol each), F3 and B3 (10 pmol each), LB and LF (20 pmol each) and 2 μL of the template. The mixture was placed at 63 °Cfor 60 min, and ddH2O was used as the control. Three replicates of each sample were run for LAMP and real-time PCR.
Real-time turbidimetry and Tris-EDTA visual reagent (TVR, patent-pending, JXD Co., China) were used to evaluate the reaction results. White precipitates of Mg2P2O7 were produced during the LAMP reaction, and thus, the turbidity of the reaction tube was monitored every 6 s by a real-time turbidimeter (La-320, Eiken Chemical Co., Japan). A curve was then drawn to judge whether the reaction was positive. The addition of the TVR indicator to the reaction system resulted in different colors. Once double-chain nucleic acids appear in the amplification system, the dye groups can bind to them and show a bright green color, indicating a positive result; a colorless solution is deemed a negative result.
The LAMP kit was obtained from Eiken Chemical Co., Japan. The LAMP reaction system (25 μL) contained 12.5 μL of 2 × buffer, 1 μL of enzyme mixture, FIP and BIP (40 pmol each), F3 and B3 (10 pmol each), LB and LF (20 pmol each) and 2 μL of the template. The mixture was placed at 63 °Cfor 60 min, and ddH2O was used as the control. Three replicates of each sample were run for LAMP and real-time PCR.
Real-time turbidimetry and Tris-EDTA visual reagent (TVR, patent-pending, JXD Co., China) were used to evaluate the reaction results. White precipitates of Mg2P2O7 were produced during the LAMP reaction, and thus, the turbidity of the reaction tube was monitored every 6 s by a real-time turbidimeter (La-320, Eiken Chemical Co., Japan). A curve was then drawn to judge whether the reaction was positive. The addition of the TVR indicator to the reaction system resulted in different colors. Once double-chain nucleic acids appear in the amplification system, the dye groups can bind to them and show a bright green color, indicating a positive result; a colorless solution is deemed a negative result.
5
LAMP Assay Optimization for EGFR
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The LAMP assays were performed using a commercially available LAMP kit (Eiken Chemical, Japan). Optimization of parameters such as temperature, time, sensitivity, and specificity was carried out using the wild-type LAMP primers and wild-type plasmid DNA construct (EGFR WT, #11011). The reaction mixtures for optimization assays were prepared as per the manufacturer&s instructions, which included 12.5μl reaction mixture containing 20 μM each of FIP and BIP and 2.5 μM of F3 and B3.
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