The detection and identification of saliva in forensic samples by RT-LAMP.

We report on a novel method for saliva identification by reverse transcription-loop-mediated isothermal amplification (RT-LAMP). In our previous report, real-time RT-LAMP was used for blood identification by using HBB detection as a model but in this advanced study, this method was refined for the identification of the more challenging body fluid of saliva. Expression of the18S rRNA gene was used as the internal control and the Statherin (STATH) gene as the saliva-specific marker. A turbidimeter was used for real-time detection of the RT-LAMP products, and confirmation was obtained that the real products were generated using: agarose gel electrophoresis, calcein fluorescence detection and/or enzymatic digestion. The specificity of the test was performed using 42 samples including 7 different body fluids, and the expression of STATH was only observed in all the saliva samples (6) with a threshold time of 39.4 ± 2.9 min. Sensitivity testing showed that RT-LAMP products for STATH were stably detected when the RNA template was not less than 6.25 ng. When the primer concentrations for STATH were two times that of 18S rRNA, saliva could be identified in the body fluid mixtures even at a ratio (saliva:semen) of 1:3 (without loop primer)/1:5 (with loop primer). A multiplex RT-LAMP was established to simultaneously amplify the 18S rRNA and STATH genes, and applied to the identification of saliva on ten non-probative cigarette butts. A positive result for saliva was obtained from all ten butts, even for those that returned a negative or ambiguous result using the amylase test. A direct RT-LAMP test is also reported where the RNA extraction step was omitted to speed the collection of data and all tests using either the simplex or multiplex RT-LAMP resulted in a positive response if saliva was present. Our data provide a simple and effective means to detect the presence of saliva.

Precise frequency measurements of iodine hyperfine transitions at 671 nm.

We report absolute frequency measurements on the a(1), a(10), and a(15) hyperfine components of the R(78) 4-6 line of (127)I(2). An external-cavity diode laser system at 671 nm is frequency-stabilized to the saturated absorption center obtained by modulation transfer spectroscopy in an iodine vapor cell. Its absolute frequency is measured by an optical frequency comb. The effect of pressure shift is investigated to obtain the absolute transition frequency at zero pressure. Our determination of the line centers reaches a precision of better than 40 kHz and will provide useful input for theoretical calculations. This frequency-stabilized laser can be used as a reference laser for the spectroscopy of lithium D lines.

Saturation spectroscopy of CO2 and frequency stabilization of an optical parametric oscillator at 2.77 µm.

We report the frequency stabilization of a CW single-frequency, singly resonant optical parametric oscillator (OPO) to the saturation absorption center of the (12)C(16)O2[10°1,02°1](II)>←00°0 P(14) line at 2.77 µm. The CO2 molecules were excited by the OPO idler wave, and the absorption signal was monitored through the fluorescence at 4.3 µm using a gold-coated longitudinal cell. The idler frequency was stabilized onto the line center by wavelength modulation method. The linewidth of the saturation dip was estimated to be 4.7 MHz, and the achieved frequency stability was 3.9 kHz (3.6×10(-11)).

High-resolution sub-Doppler Lamb dips of the ν2 fundamental band of H3(+).

The high-resolution sub-Doppler Lamb dips of the ν2 fundamental band transitions of H3(+) have been observed using an extended negative glow discharge tube as an ion source and a periodically poled lithium niobate optical parametric oscillator as a radiation source. The absolute frequency of the R(1,0) transition was measured to be 81,720,371.550 MHz with an accuracy of 250 kHz using an optical frequency comb. In addition, we have investigated the linewidth of the Lamb-dip signal of the R(3,0) transition systematically and obtained its pressure-broadening parameter, which may shed some light on the reaction of H3(+) with H2. This is the first observation of the infrared saturated spectrum and the first determination of the pressure-broadening parameter of the ro-vibrational transitions of a molecular ion.